Dry powder formulations for inhalation

ABSTRACT

A respirable dry powder can include acetylsalicylic acid in particles having a mass median aerodynamic diameter (MMAD) within a range of about 0.5 μm to about 10 μm. The respirable dry powder may contain a pharmaceutically acceptable excipient, such as an amino acid (e.g., Leucine), in an amount ranging from about 0.1% (w/w) to about 40% (w/w) of the particles.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.15/613,123, filed Jun. 2, 2017, which claims priority to U.S.Provisional Application No. 62/345,123, filed Jun. 3, 2016, the contentsof which are incorporated herein by reference in their entirety.

FIELD

The subject technology relates generally to pulmonary delivery of drypowder formulations of nonsteroidal anti-inflammatories (NSAIDs), suchas aspirin or acetylsalicylic acid. The subject technology also relatesgenerally to apparatuses and methods for delivery of substances, e.g.,medication, to the lungs by inhalation for treating disease, such asischemic or thromboembolic events, including cardiovascular disease.

BACKGROUND

Pulmonary delivery of therapeutic agents offers several advantages overother modes of delivery. These advantages include rapid onset, theconvenience of patient self-administration, the potential for reduceddrug side-effects, ease of delivery by inhalation, the elimination ofneedles, and the like. Inhalation therapy is capable of providing a drugdelivery system that is easy to use in an inpatient or outpatientsetting, results in very rapid onset of drug action, and producesminimal side effects.

SUMMARY

The present dry powder composition comprises at least one NSAID, such asacetylsalicylic acid, as the active ingredient. The dry powdercomposition contains dry particles that comprise acetylsalicylic acid ora pharmaceutically acceptable salt thereof. The dry particles can berespirable. The dry particles may have a mass median aerodynamicdiameter (MMAD) ranging from about 0.5 μm to about 10 μm, from about 1μm to about 10 μm, from about 1 μm to about 5 μm, from about 3 μm toabout 4 μm, from about 2 μm to about 5 μm, or about 20 μm or less. Thedry particles may vary in size, e.g., with a geometric diameter (VMGD)between 0.5 μm and 30 μm.

The composition may further comprise an amino acid, a polypeptide, orcombinations thereof. The amino acid and/or polypeptide may bewater-soluble. The amino acid may be an L-amino acid, a D-amino acid ora combination thereof. The amino acid may be Leucine, Alanine, Arginine,Asparagine, Aspartic acid, Cysteine, Glutamic acid, Glutamine, Glycine,Histidine, Isoleucine, Lysine, Methionine, Phenylalanine, Proline,Serine, Threonine, Tryptophan, Tyrosine, Valine, or combinations orvariants thereof.

The amino acid (e.g., leucine) and/or polypeptide may be in an amountranging from about 0.1% (w/w) to about 10% (w/w), from about 0.1% (w/w)to about 40% (w/w), from about 1% to about 30% (w/w), from about 0.5% toabout 20% (w/w), from about 0% to about 99% (w/w), from about 0.01% toabout 80% (w/w), from about 0.05% to about 70% (w/w), from about 0.1% toabout 60% (w/w), from about 0.1% to about 50% (w/w), from about 0.1% toabout 40% (w/w), from about 0.1% to about 30% (w/w), from about 0.1% toabout 20% (w/w), from about 0.05% to about 8% (w/w), from about 0.1% toabout 6% (w/w), from about 5% to about 10% (w/w), from about 3% to about8% (w/w), from about 2% to about 6% (w/w), from about 0.1% to about 5%(w/w), from about 0.1% to about 4% (w/w), from about 0.1% to about 3%(w/w), from about 0.1% to about 2% (w/w), from about 0.1% to about 1%(w/w), from about 1% to about 6% (w/w), from about 1% to about 5% (w/w),from about 1% to about 4% (w/w), or from about 1% to about 3% (w/w),about 0.1% (w/w), about 5% (w/w), about 4% (w/w), about 13% (w/w), orabout 15% (w/w), of the composition.

The composition may further comprise a pharmaceutically acceptableexcipient, such as one or more phospholipids, in an amount ranging fromabout 0.1% (w/w) to about 10% (w/w), from about 0% to about 99% (w/w),from about 0.01% to about 80% (w/w), from about 0.05% to about 70%(w/w), from about 0.1% to about 60% (w/w), from about 0.1% to about 50%(w/w), from about 0.1% to about 40% (w/w), from about 0.1% to about 30%(w/w), from about 0.1% to about 20% (w/w), from about 0.05% to about 8%(w/w), from about 0.1% to about 6% (w/w), from about 5% to about 10%(w/w), from about 3% to about 8% (w/w), from about 2% to about 6% (w/w),from about 0.1% to about 5% (w/w), from about 0.1% to about 4% (w/w),from about 0.1% to about 3% (w/w), from about 0.1% to about 2% (w/w),from about 0.1% to about 1% (w/w), from about 1% to about 6% (w/w), fromabout 1% to about 5% (w/w), from about 1% to about 4% (w/w), or fromabout 1% to about 3% (w/w), about 0.1% (w/w), about 5% (w/w), about 3%(w/w), or about 10% (w/w), of the composition.

Non-limiting examples of the phospholipids include dipalmitoylphosphatidylcholine (DPPC), distearoyl phosphatidylcholine (DSPC), soylecithin, or a combination thereof. In one embodiment, the weightpercentage of the DSPC is 5% (w/w) of the composition. In anotherembodiment, soy lecithin is 0.1% (w/w) of the composition.

The dry particles may be coated with a pharmaceutically acceptableexcipient, such as one or more phospholipids, amino acids, polypeptides,and any combinations thereof.

Acetylsalicylic acid, or a pharmaceutically acceptable salt thereof, maybe in an amount greater than 20% (w/w), greater than 30% (w/w), greaterthan 40% (w/w), greater than 50% (w/w), greater than 60% (w/w), greaterthan 70% (w/w), greater than 80% (w/w), greater than 90% (w/w), of thecomposition.

The MMAD of the dry particles may vary less than about 10%, less thanabout 8%, less than about 6%, less than about 4%, or less than about 2%,after the dry powder composition is stored at 30° C. at 65% relativehumidity for about 4 weeks, after the dry powder composition is storedat 50° C. at 75% relative humidity for about 2 weeks, or after the drypowder composition is stored at 50° C. for about 5 days.

The particles may have an MMAD size distribution where the particlesexhibit: (i) a DV90 less than about 20 μm, a DV50 less than about 7 μm,and a DV10 less than about 2 μm; (ii) a DV90 less than about 10 μm, aDV50 less than about 4 μm, and a DV10 less than about 1 μm; (iii) a DV90less than about 6 μm, a DV50 less than about 3 μm, and a DV10 less thanabout 1 μm; (iv) a DV50 less than about 5 μm, and a DV10 less than about2 μm; or (v) a DV50 ranging from about 2.5 μm to about 4 μm, and a DV10ranging from about 0.8 μm to about 1.5 m.

The DV90, DV50 and/or DV10 of the dry particles may vary less than about10%, less than about 8%, less than about 6%, less than about 4%, or lessthan about 2%, after the dry powder composition is stored at 30° C. at65% relative humidity for about 4 weeks.

The subject technology also provides a drug delivery system for treating(including prophylactic treatment or reducing the risk of) acardiovascular disease (such as thrombosis), the system comprising thepresent composition. Acetylsalicylic acid may be present at a doseranging from about 2 mg to about 80 mg, from about 5 mg to about 80 mg,from about 5 mg to about 60 mg, from about 5 mg to about 50 mg, fromabout 5 mg to about 40 mg, or from about 10 mg to about 40 mg.

In certain embodiments, the drug delivery system contains: atherapeutically effective dose of an NSAID (such as acetylsalicylicacid) in dry powder form; a dry powder inhaler, the dry powder inhalercomprising a mouthpiece, a reservoir for receiving the dose of the NSAID(such as acetylsalicylic acid), and an actuation member for makingavailable the dose of the acetylsalicylic acid for inhalation by thepatient through the mouthpiece. A single inhaled dose of the NSAID (suchas acetylsalicylic acid) may be about 40 mg or less, or 30 mg or less.The dose of acetylsalicylic acid may be present at amounts ranging fromabout 5 to about 40 mg.

The formulation may further comprise clopidogrel.

In one embodiment, the pharmaceutically acceptable excipient is DSPC,the respirable dry powders substantially comprise dry particles having aMMAD ranging from about 3 to about 4 μm. The mass percent of stages inan NGI testing apparatus of the respirable powder yields may be at,stage 1 about 10% to about 13%, stage 2, about 20% to about 23%, stage3, about 13% to about 15%, and stage 4, about 5% to about 6% and fineparticle fraction ranges from about 45% to about 55%.

In another embodiment, the pharmaceutically acceptable excipient is soylecithin, the respirable dry powders substantially comprise dryparticles having a MMAD ranging from about 2.0 to about 3.0 μm. The masspercent of stages in an NGI testing apparatus of the respirable powderyields are at, stage 1 about 5% to about 10%, stage 2, about 10% toabout 18%, stage 3, about 15% to about 20%, and stage 4, about 10% toabout 15% and fine particle fraction ranges from about 50% to about 70%.

The composition may further comprise an excipient such as sodium laurylsulfate (SLS), lactose, starch, cellulose, leucine, sodium citrate,maltodextrin, mannitol or a combination thereof.

In one embodiment, the particles may have a size distribution where 90%of the formulation comprises particles with an MMAD of about 6 μm orless, 50% of the formulation comprises particles having an MMAD of about3 μm or less, and 10% of the formulation comprises particles having anMMAD of about 1 μm or less.

The respirable dry powder compositions can include a pharmaceuticallyacceptable excipient, such as leucine, sodium citrate, maltodextrin ormannitol, which may be present in an amount of about 5% to about 90% orby weight.

In one embodiment, the NSAID, such as acetylsalicylic acid, is providedin a dry powder formulation comprising a mixture of particles of varioussizes, for example, a mixture of (i) particles having a mean geometricdiameter (VMGD) and/or mass median aerodynamic diameter (MMAD) of about5 μm or less, and (ii) particles having a mean geometric diameter (VMGD)and/or mass median aerodynamic diameter (MMAD) of 15 μm or greater. Inone embodiment, the composition may include a pharmaceuticallyacceptable excipient. In another embodiment, the composition is free orsubstantially free of excipient. In certain embodiments, the compositionis free or substantially free of anti-aggregation excipient.

In another embodiment, the NSAID, such as acetylsalicylic acid,comprises dry particles having a mass median aerodynamic diameter (MMAD)within a range of about 0.5 μm to about 10 μm, wherein the dry powderfurther comprises one or more phospholipids in an amount ranging fromabout 0.1% (w/w) to about 10% (w/w) of the dry particles. The particlesmay have an MMAD size distribution where the particles exhibit: (i) aDV90 less than about 20 μm, a DV50 less than about 7 μm, and a DV10 lessthan about 2 m; (ii) a DV90 less than about 10 μm, a DV50 less thanabout 4 μm, and a DV10 less than about 1 m; or (iii) a DV90 less thanabout 6 μm, a DV50 less than about 3 μm, and a DV10 less than about 1μm.

In another embodiment, the NSAID, such as acetylsalicylic acid,comprises dry particles having a mass median aerodynamic diameter (MMAD)within a range of about 0.5 μm to about 10.0 μm, about 2.0 μm to about5.0 μm, about 3.0 μm to about 4.0 μm, wherein the dry powder furthercomprises one or more amino acids (e.g., leucine) and/or polypeptides inan amount ranging from about 0.1% (w/w) to about 40% (w/w), about 0.1%(w/w) to about 30% (w/w), about 0.1% (w/w) to about 20% (w/w), about 2%(w/w) to about 20% (w/w), about 4% (w/w) to about 15% (w/w), about 4%(w/w), about 5% (w/w), about 13% (w/w), or about 15% (w/w) of the dryparticles. The particles may have an MMAD size distribution where theparticles exhibit: (i) a DV90 less than about 10 μm, a DV50 less thanabout 5 μm, and a DV10 less than about 2 m; (ii) a DV90 less than about5 μm, a DV50 less than about 2.1 μm, and a DV10 less than about 1 m; or(iii) a DV90 less than about 4 μm, a DV50 less than about 2.0 μm, and aDV10 less than about 1 μm.

The present application also provides for methods in therapy (e.g.,treatment, prophylaxis, or diagnosis). The present composition may beused for treatment (including prophylactic treatment, such as preventionor reducing the risk) of a cardiovascular disease (such thrombosis), andin the manufacture of a medicament for the treatment, prophylaxis ordiagnosis of a cardiovascular or thromboembolic disease (such asthrombosis). In certain embodiments, the present application providesfor a method of treating an ischemic event, reducing the risk of athromboembolic event or treating thrombosis. The method containsadministrating to a subject in need thereof a therapeutically effectivedose of the present dry powder composition.

The thromboembolic event may be a myocardial infarction, unstableangina, or a stroke. The thromboembolic event may be a transientischemic attack.

The thromboembolic event may be treated within about 5, 10 or 15 minutesof onset of an ischemic event.

BRIEF DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawings executedin color. Copies of this patent or patent application publication withcolor drawing(s) will be provided by the Office upon request and paymentof the necessary fee.

The accompanying drawings, which are included to provide furtherunderstanding of the subject technology and are incorporated in andconstitute a part of this specification, illustrate aspects of thesubject technology and together with the description serve to explainthe principles of the subject technology.

FIG. 1 shows laser diffraction data of Formulation 3727.

FIG. 2 shows laser diffraction data of Formulation 3734.

FIG. 3 shows DSC thermograms of raw, micronized uncoated and spray-driedDSPC/acetylsalicylic acid particles.

FIG. 4 shows TGA of micronized uncoated and spray-driedDSPC/acetylsalicylic acid particles.

FIGS. 5A and 5B show the particle size distribution of spray-driedDSPC/acetylsalicylic acid particles based on NGI analysis. FIG. 5A: 2capsules were delivered to the NGI; FIG. 5B: 1 capsule was delivered tothe NGI.

FIGS. 6A and 6B show the particle size distribution of spray-dried soylecithin/acetylsalicylic acid particles based on NGI analysis. FIG. 6A:2 capsules were delivered to the NGI; FIG. 6B: 1 capsule was deliveredto the NGI.

FIGS. 7A and 7B show laser diffraction data (FIG. 7A) and morphology(FIG. 7B) of milled control (BREC-1511-024, 100% jet-milled ASA), 100%ASA (BREC-1511-038A, spray dried from hexane), and 99.9/0.1 ASA/Lecithin(BREC-1511-038B, spray dried from hexane) formulations.

FIG. 8 shows laser diffraction data of 100% jet-milled ASA(BREC-1511-024), BREC-1511-038A (spray dried from hexane, 100% ASA),spray dried from hexane 99.9/0.1 ASA/Lecithin (BREC-1511-038B) over theperiod of four weeks (time 0, week 1, week 2, and week 4) at 30° C. and65% relative humidity (RH).

FIG. 9 shows the particle size distribution of BREC1511-024 particlesbased on NGI analysis (week 4, at 30° C. and 65% RH).

FIG. 10 shows the particle size distribution of BREC1511-038A particlesbased on NGI analysis (week 4, at 30° C. and 65% RH).

FIG. 11 shows the particle size distribution of BREC1511-038B particlesbased on NGI analysis (week 4, at 30° C. and 65% RH).

FIG. 12 shows particle morphology of BREC1511-024, BREC1511-038A, andBREC1511-038B (99.9/0.1 ASA/Lecithin) formulations (at initial timepoint and 2 weeks, at 50° C./75% RH).

FIG. 13 is a summary of RP-HPLC results of BREC1511-024, BREC1511-038A,and BREC1511-038B after 2 weeks at 50° C./75% RH.

FIG. 14 is a graph of particle size distribution of BREC1511-024,BREC1511-038A, and BREC1511-038B after 2 weeks at 50° C./75% RH.

FIG. 15 shows the particle size distribution of BREC1511-024 particlesbased on NGI analysis (after 2 weeks at 50° C./75% RH).

FIG. 16 shows the particle size distribution of BREC1511-038A particlesbased on NGI analysis (after 2 weeks at 50° C./75% RH).

FIG. 17 shows the particle size distribution of BREC1511-038B particlesbased on NGI analysis (after 2 weeks at 50° C./75% RH).

FIG. 18 shows laser diffraction data for (BREC-1511-052A), Spray Dried100% Jet Milled ASA (High Flow)(BREC-1511-052B), Spray Dried 100% JetMilled ASA (High Flow, High Solids)(BREC-1511-052C), Spray Dried 100%Jet Milled ASA (High Flow, High Solids, High Tout)(BREC-1511-052D), andSpray Dried 99.9/0.1 Jet Milled ASA/Lecithin (HighFlow)(BREC-1511-052E).

FIG. 19 shows particle morphology of BREC-1511-052A, BREC-1511-052B,BREC-1511-052C, BREC-1511-052D, and BREC-1511-052E (images were obtainedusing SEM).

FIG. 20 shows powder characteristics for each batch (BREC-1511-052A,BREC-1511-052B, BREC-1511-052C, BREC-1511-052D, and BREC-1511-052E).

FIG. 21 shows ASA and L-leucine absolute solubility.

FIG. 22 shows a scheme for studying ASA and L-leucine absolutesolubility.

FIG. 23 shows a scheme for spray dry process optimization.

FIG. 24A—show laser diffraction data of various examples of 96/4Aspirin/Leucine (BREC-1511-178A) and 87/13 Aspirin/L-Leucine(BREC-1511-178B) formulations as disclosed herein, respectively:

FIG. 24A: laser diffraction data of examples of 96/4 Aspirin/L-Leucine(BREC-1511-178A) and 87/13 Aspirin/L-Leucine (BREC-1511-178B)formulations as disclosed herein.

FIG. 24B: laser diffraction data of an example of 96/4 Aspirin/L-Leucine(BREC-1511-178A) formulation as disclosed herein before and afterstorage at various conditions for two months.

FIG. 24C: laser diffraction data of an example of 87/13Aspirin/L-Leucine (BREC-1511-178B) formulation as disclosed hereinbefore and after storage at various conditions for two months.

FIG. 24D: D(v 0.9) of an example of 96/4 Aspirin/L-Leucine(BREC-1511-178A) formulation as disclosed herein before and afterstorage at various conditions for two weeks, one month or two months.

FIG. 24E: D(v 0.9) of an example of 87/13 Aspirin/L-Leucine(BREC-1511-178B) formulation as disclosed herein before and afterstorage at various conditions for two weeks, one month or two months.

FIG. 24F: laser diffraction data of an example of 95/5 Aspirin/L-Leucine(BREC-1688-046) formulation as disclosed herein before and after storageat various conditions for one month.

FIG. 24G: laser diffraction data of an example of 85/15Aspirin/L-Leucine (BREC-1688-036) formulation as disclosed herein beforeand after storage at various conditions for one month.

FIG. 24H: laser diffraction data of an example of 95/5 Aspirin/L-Leucine(BREC-1688-046) formulation as disclosed herein before and after storageat various conditions for six months.

FIG. 24I: laser diffraction data of an example of 85/15Aspirin/L-Leucine (BREC-1688-036) formulation as disclosed herein beforeand after storage at various conditions for six months.

FIG. 24J: D(v 0.9) of an example of 95/5 Aspirin/L-Leucine(BREC-1688-046) formulation as disclosed herein before and after storageat various conditions for one or six months.

FIG. 24K: D(v 0.9) of an example of 85/15 Aspirin/L-leucine(BREC-1688-036) formulation as disclosed herein before and after storageat various conditions for one or six months.

FIGS. 25A-25G show morphology of examples of 96/4 Aspirin/L-leucine(BREC-1511-178A), 87/13 Aspirin/L-leucine (BREC-1511-178B), 95/5Aspirin/L-leucine (BREC-1688-046), and 85/15 Aspirin/L-leucine(BREC-1688-036) formulations as disclosed herein. The images wereobtained using SEM (scanning electron microscopy):

FIG. 25A: SEM images of examples of 96/4 Aspirin/L-leucine(BREC-1511-178A) and 87/13 Aspirin/L-leucine (BREC-1511-178B)formulations as disclosed herein.

FIG. 25B: SEM images of an example of 96/4 Aspirin/L-leucine(BREC-1511-178A) formulation as disclosed herein before and afterstorage at various conditions for two months.

FIG. 25C: SEM images of an example of 87/13 Aspirin/L-leucine(BREC-1511-178B) formulation as disclosed herein before and afterstorage at various conditions for two months.

FIG. 25D: SEM images of an example of 95/5 Aspirin/L-leucine(BREC-1688-046) formulation as disclosed herein before and after storageat various conditions for one month.

FIG. 25E: SEM images of an example of 85/15 Aspirin/L-leucine(BREC-1688-036) formulation as disclosed herein before and after storageat various conditions for one month.

FIG. 25F: SEM images of an example of 95/5 Aspirin/L-leucine(BREC-1688-046) formulation as disclosed herein before and after storageat various conditions for six months.

FIG. 25G: SEM images of an example of 85/15 Aspirin/L-leucine(BREC-1688-036) formulation as disclosed herein before and after storageat various conditions for six months.

FIGS. 26A-26E show aerosol profile studies by NGI of examples of theASA/L-leucine formulations as disclosed herein:

FIG. 26A: Aerosol profile studies by NGI of examples of 96/4Aspirin/L-leucine (BREC-1511-178A) and 87/13 Aspirin/L-leucine(BREC-1511-178B) formulations as disclosed herein.

FIG. 26B: Aerosol profile studies by NGI of an example of 95/5Aspirin/L-leucine (BREC-1688-046) formulation as disclosed herein.

FIG. 26C: Aerosol profile studies by NGI of an example of 95/5Aspirin/L-leucine (BREC-1688-046) formulation as disclosed herein.

FIG. 26D: Aerosol profile studies by NGI of an example of 85/15Aspirin/L-leucine (BREC-1688-036) formulation as disclosed herein.

FIG. 26E: Aerosol profile studies by NGI of an example of 85/15Aspirin/L-leucine (BREC-1688-036) formulation as disclosed herein.

FIG. 27 shows XRPD analysis of various examples of ASA/L-leucineformulations as disclosed herein.

FIGS. 28A-FIG. 28E shows ASA purity or SA impurity measured by RP-HPLCof various examples of ASA/L-leucine formulations as disclosed herein,before and after exposed to various storage conditions:

FIG. 28A: RP-HPLC chromatogram of an example of 96/4 Aspirin/L-leucine(BREC-1511-178A) formulation as disclosed herein before and afterstorage at various conditions for two months.

FIG. 28B: RP-HPLC chromatogram of an example of 87/13 Aspirin/L-leucine(BREC-1511-178B) formulation as disclosed herein before and afterstorage at various conditions for two months.

FIG. 28C: SA content measured by RP-HPLC of an example of 96/4Aspirin/L-leucine (BREC-1511-178A) formulation as disclosed hereinbefore and after storage at various conditions for two months.

FIG. 28D: SA content measured by RP-HPLC of an example of 87/13Aspirin/L-leucine (BREC-1511-178B) formulation as disclosed hereinbefore and after storage at various conditions for two months.

FIG. 28E: RP-HPLC chromatogram of an example of 95/5 Aspirin/L-leucine(BREC-1688-046) formulation as disclosed herein before and after storageat various conditions for one or six months.

FIG. 28F: RP-HPLC chromatogram of an example of 85/15 Aspirin/L-leucine(BREC-1688-036) formulation as disclosed herein before and after storageat various conditions for one or six months.

DETAILED DESCRIPTION

As set forth in the Examples section below, various aspirin-leucine drypowder compositions were found to have unexpected high chemical andparticle stabilities after being stored at various temperature andhumidity conditions for various period of time. Four aspirin-leucine drypowder compositions with various formulations were prepared byspray-drying method with two solvent mixtures of EtOH and water. RP-HPLCanalysis showed all aspirin-leucine dry powder compositions maintainedan aspirin purity of higher than 99% after stored at 4° C. at 45% RH orlower for about two months, at 25° C. at 60% RH for about two months,stored at 30° C. at 65% RH for about one month, or stored at 40° C. at75% RH for about two months. After left for about six months at 25° C.at 60% RH, 30° C. at 65% RH, or 40° C. at 75% RH, the test formulationsshowed an aspirin purity of higher than about 97%. SEM images of variousaspirin-leucine dry powder compositions tested as described in Example12 showed rod-like crystals with rod-like crystals with small and roughspheres. Such morphology was maintained after the aspirin-leucine drypowder compositions were stored at 4° C. at 45% RH or lower for about 2weeks, about one month, or about two months, stored at 25° C. at 60% RHfor about 2 weeks, about one month, about two months, or about sixmonths, stored at 30° C. at 65% RH for about 2 weeks, about one month,about two months, or about six months, stored at 40° C. at 75% RH forabout 2 weeks, about one month, about two months, or about six months.Particle size of the test aspirin-leucine dry powder formulation alsoshowed insignificant changes after stored under the test conditions. Oneor more particle size parameters, e.g., MMAD, D (v 0.1), D (v0.5),D(v0.9), D[3,2], D[4,3] and span of the dry particles, showed a changeof about 10% or lower, about 5% or lower, or about 2.5% or lower, afterstorage at 4° C., 25° C./60% RH, 30° C./65% RH, or 40° C./75% RH for onemonth, two months, or six months.

Accordingly, a dry powder composition containing acetylsalicylic acid isprovided herein with desired chemical and particle stability. The dryparticles of the dry powder may have a mass median aerodynamic diameter(MMAD) within a range of about 0.5 μm to about 10 μm. The respirable drypowder may contain a pharmaceutically acceptable excipient, such as oneor more phospholipids, amino acids (e.g., leucine), and/or apolypeptides, in an amount ranging from about 0.1% (w/w) to about 10%(w/w), from about 0.1% (w/w) to about 40% (w/w), from about 1% to about30% (w/w), from about 0.5% to about 20% (w/w), from about 0% to about99% (w/w), from about 0.01% to about 80% (w/w), from about 0.05% toabout 70% (w/w), from about 0.1% to about 60% (w/w), from about 0.1% toabout 50% (w/w), from about 0.1% to about 40% (w/w), from about 0.1% toabout 30% (w/w), from about 0.1% to about 20% (w/w), from about 0.05% toabout 8% (w/w), from about 0.1% to about 6% (w/w), from about 5% toabout 10% (w/w), from about 3% to about 8% (w/w), from about 2% to about6% (w/w), from about 0.1% to about 5% (w/w), from about 0.1% to about 4%(w/w), from about 0.1% to about 3% (w/w), from about 0.1% to about 2%(w/w), from about 0.1% to about 1% (w/w), from about 1% to about 6%(w/w), from about 1% to about 5% (w/w), from about 1% to about 4% (w/w),from about 1% to about 3% (w/w), about 0.1%, about 5% (w/w), about 4%(w/w), about 3%, about 10% (w/w), about 13% (w/w), or about 15% (w/w),of the composition of the particles.

The dry particles of the present composition show stability. In certainembodiments, the MMAD of the dry particles of the present compositionvaries less than about 30%, less than about 25%, less than about 20%,less than about 15%, less than about 10%, less than about 9%, less thanabout 8%, less than about 7%, less than about 6%, less than about 5%,less than about 4%, less than about 3%, less than about 2.5%, less thanabout 2%, or less than about 1%, after the composition is stored at 30°C. at 65% relative humidity (RH) for about 4 weeks, stored at 50° C. at75% relative humidity for about 2 weeks, stored at 4° C. at 45% RH orlower for about 2 weeks, about one month, or about two months, stored at25° C. at 60% RH for about 2 weeks, about one month, about two months,or about six months, stored at 30° C. at 65% RH for about 2 weeks, aboutone month, about two months, or about six months, stored at 40° C. at75% RH for about 2 weeks, about one month, about two months, or aboutsix months. In certain embodiments, the DV90, DV50 and/or DV10 of thedry particles of the present composition vary (varies) less than about30%, less than about 25%, less than about 20%, less than about 15%, lessthan about 10%, less than about 9%, less than about 8%, less than about7%, less than about 6%, less than about 5%, less than about 4%, lessthan about 3%, less than about 2.5%, less than about 2%, or less thanabout 1%, after the composition is stored at 30° C. at 65% relativehumidity for about 4 weeks, or stored at 50° C. at 75% relative humidityfor about 2 weeks, stored at 4° C. at 45% RH or lower for about 2 weeks,about one month, or about two months, stored at 25° C. at 60% RH forabout 2 weeks, about one month, about two months, or about six months,stored at 30° C. at 65% RH for about 2 weeks, about one month, about twomonths, or about six months, stored at 40° C. at 75% RH for about 2weeks, about one month, about two months, or about six months.

1. Introduction

Thromboembolic Symptoms and Events

A thromboembolic event, such as myocardial infarction, deep venousthrombosis, pulmonary embolism, thrombotic stroke, etc., can presentwith a group of symptoms that allow a patient or clinician to provide aninitial therapy or treatment for the event, i.e., immediately, or within1, 5, 10 or 15 minutes of onset of the thromboembolic event. In certainsituations, an 81 mg, low dose, or “baby” acetylsalicylic acid or aregular acetylsalicylic acid (330 mg) may be orally administered inorder to provide an initial treatment for the patient. However, oraladministration may not act as quickly as necessary to provide asufficient therapeutic effect and therefore, may lead to a lesspreferred outcome. Thus, the pulmonary drug delivery system and relatedmethods or the methods disclosed herein provide for an accelerated andmore efficient pathway and treatment for reducing the risk of athromboembolic event and/or providing treatment for a thromboembolicevent. For example, certain embodiments provide systems and methods ofadministering a non-steroidal anti-inflammatory drug (NSAID) byinhalation, such as by a dry powder inhaler (DPI) or a metered doseinhaler (MDI).

Dry Powder Inhaler Technology

Delivery of a drug by inhalation in the early stages of an emergencysituation can provide a fast-acting, effective form of preliminarytreatment of certain medical conditions. For example, in one embodiment,upon receiving a complaint of a symptom of a serious thromboembolicevent, a patient can be administered, by DPI, a therapeutic amount of aNSAID. The NSAID can address problems associated with or provide aninitial treatment for the medical condition.

However, dry powder inhalation of drugs has generally been limited todosages of less than a milligram. Recent developments in particleengineering, in particular the development of PulmoSphere® technology,have enabled the delivery of a larger amount of dry powder to the lungsin a single actuation. See David E. Geller, M.D., et al., Development ofan inhaled dry-powder formulation of tobramycin using Pulmosphere™technology, J Aerosol Med Pulm Drug Deliv. 2011 August; 24(4), pp.175-82. In this publication, a dose of 112 mg tobramycin (in fourcapsules) was effectively delivered via PulmoSpheres®.

To date, there has been no single dose use of acetylsalicylic acid bydry powder inhaler to replace the traditional daily use of a NSAID (suchas a baby acetylsalicylic acid) or emergency use of a NSAID aspreventative care for symptoms of a thromboembolic event. Accordingly,in one embodiment, the methods provide for administering a NSAID by drypowder inhalation in an amount less than the dosage of a babyacetylsalicylic acid (e.g., less than 81 mg).

Therefore, a method for treating disease, e.g., by reducing the risk ofa thromboembolic event, comprises administering a NSAID, such as asalicylate, by a DPI or MDI. For example, the method can compriseadministering acetylsalicylic acid by a DPI or MDI. The administereddose (e.g., a single dose) can be less than 25 mg of acetylsalicylicacid. Further, in various embodiments, the administered dosage (e.g., asingle dose) can be less than 20 mg of acetylsalicylic acid. Theadministered dosage (e.g., a single dose) can be less than 80 mg, lessthan 70 mg, less than 60 mg, less than 50 mg, less than 40 mg, less than30 mg, less than 20 mg, less than 15 mg of acetylsalicylic acid, lessthan 12 mg of acetylsalicylic acid, less than 10 mg of acetylsalicylicacid, less than 8 mg of acetylsalicylic acid, less than 5 mg ofacetylsalicylic acid, or less than 2 mg of acetylsalicylic acid.

In other embodiments, the dosage of acetylsalicylic acid (e.g., a singledose) can be from about 2 mg to about 30 mg, about 4 mg to about 25 mgof acetylsalicylic acid, about 6 mg to about 20 mg of acetylsalicylicacid, about 8 mg to about 15 mg of acetylsalicylic acid, about 10 mg toabout 13 mg of acetylsalicylic acid, about 1 mg, about 2 mg, about 3 mg,about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg,about 10 mg, about 11 mg, about 12 mg, about 13 mg, about 14 mg, about15 mg, about 16 mg, about 17 mg, about 18 mg, about 19 mg, or about 20mg of acetylsalicylic acid. Such dosages can provide a bioequivalentdosage when compared to typical dosages of about 81 mg to about 325 mg,while demonstrating few negative side effects.

Thus, a NSAID, such as acetylsalicylic acid, can be administered by DPIor MDI in a single dose or multiple dose that delivers much less than atraditional oral dose of acetylsalicylic acid, which can provide anequivalent treatment while tending to avoid the negative side effectsassociated with certain NSAIDs, such as acetylsalicylic acid. Further,systems (devices) of administering such treatments are also provided.

The NSAID, in particular acetylsalicylic acid, can be formulated toinclude pharmaceutically acceptable excipients that are effective toimprove aerodynamic performance, bioavailability and/or pharmacokineticsas compared to prior art methods of administration.

The DPI or MDI apparatus can have a mouthpiece and an actuation memberfor making available the NSAID for inhalation by a patient to reduce therisk of the thromboembolic event.

For example, a method of reducing the risk of a thromboembolic event isprovided and can comprise administering a dose of a non-steroidalanti-inflammatory drug by a dry powder inhaler. The dose can beeffective to reduce a risk of a thromboembolic event in a patient. Thedry powder inhaler can have a mouthpiece and an actuation member formaking available the dose of the non-steroidal anti-inflammatory drugfor inhalation by the patient to reduce the risk of the thromboembolicevent.

A drug delivery system can also be provided for treating a disease, forexample, by reducing the risk of a thromboembolic (ischemic) event. Thesystem can comprise a dose of a non-steroidal anti-inflammatory drug inpowder form. The dose can be effective to reduce the risk of athromboembolic event in a patient. The system can also comprise a drypowder inhaler. The dry powder inhaler can have a mouthpiece, areservoir for receiving the dose of the non-steroidal anti-inflammatorydrug, and an actuation member for making available the dose of thenon-steroidal anti-inflammatory drug for inhalation by the patientthrough the mouthpiece.

The thromboembolic event may be a myocardial infarction, deep venousthrombosis, pulmonary embolism, or thrombotic stroke. The dose of theNSAID drug can be administered as a preliminary treatment in response toa symptom of a thromboembolic event. The NSAID may be acetylsalicylicacid and may be administered in a single dose or in multiple doses,e.g., 2, 3, 4, 5, 6, 7, 8, 9 or 10 or greater doses.

2. Definitions

The present dry particles or dry powders are suitable for delivery tothe respiratory tract (e.g., pulmonary delivery) in a subject byinhalation. The dry particles may have a mass median aerodynamicdiameter (MMAD) of less than about 20 μm, less than about 15 μm, lessthan about 10 μm, less than about 5 μm, from about 1 μm to about 10 μm,from about 1 μm to about 5 μm, from about 1 μm to about 3 μm, from about1.7 μm to about 2.7 μm, or less.

The term “dispersible” describes the characteristic of a dry powder ordry particles to be dispelled into a respirable aerosol. Dispersibilityof a dry powder or dry particles is expressed herein as the quotient ofthe volume median geometric diameter (VMGD) measured at a dispersion(i.e., regulator) pressure of 1 bar divided by the VMGD measured at adispersion (i.e., regulator) pressure of 4 bar, or VMGD at 0.5 bardivided by the VMGD at 4 bar as measured by HELOS/RODOS. These quotientsare referred to herein as “¼ bar,” and “0.5/4 bar,” respectively, anddispersibility correlates with a low quotient. For example, ¼ bar refersto the VMGD of respirable dry particles or powders emitted from theorifice of a RODOS dry powder disperser (or equivalent technique) atabout 1 bar, as measured by a HELOS or other laser diffraction system,divided the VMGD of the same respirable dry particles or powdersmeasured at 4 bar by HELOS/RODOS. Thus, a highly dispersible dry powderor dry particles will have a ¼ bar or 0.5/4 bar ratio that is close to1.0. Highly dispersible powders have a low tendency to agglomerate,aggregate or clump together and/or, if agglomerated, aggregated orclumped together, are easily dispersed or de-agglomerated as they emitfrom an inhaler and are breathed in by the subject. Dispersibility canalso be assessed by measuring the size emitted from an inhaler as afunction of flow rate.

The term “emitted dose” or “ED” refers to an indication of the deliveryof a drug formulation from a suitable inhaler device after a firing ordispersion event. More specifically, for dry powder formulations, the EDis a measure of the percentage of powder that is drawn out of a unitdose package and that exits the mouthpiece of an inhaler device. The EDis defined as the ratio of the dose delivered by an inhaler device tothe nominal dose (i.e., the mass of powder per unit dose placed into asuitable inhaler device prior to firing). The ED is anexperimentally-measured parameter, and can be determined using themethod of USP Section 601 Aerosols, Metered-Dose Inhalers and Dry PowderInhalers, Delivered-Dose Uniformity, Sampling the Delivered Dose fromDry Powder Inhalers, United States Pharmacopeia Convention, Rockville,Md., 13th Revision, 222-225, 2007. This method utilizes an in vitrodevice set-up to mimic patient dosing.

The terms “FPF (<5.6),” “FPF (<5.6 μm),” and “fine particle fraction ofless than 5.6 μm” refer to the fraction of a sample of dry particlesthat have an aerodynamic diameter of less than a certain size, e.g., 5.6μm. For example, FPF (<5.6) can be determined by dividing the mass ofrespirable dry particles deposited on the stage one and on thecollection filter of a two-stage collapsed Andersen Cascade Impactor(ACI) by the mass of respirable dry particles weighed into a capsule fordelivery to the instrument. This parameter may also be identified as“FPF_TD (<5.6),” where TD means total dose. A similar measurement can beconducted using an eight-stage ACI. The eight-stage ACI cutoffs aredifferent at the standard 60 L/min flow rate, but the FPF_TD (<5.6) canbe extrapolated from the eight-stage complete data set. The eight-stageACI result can also be calculated by the USP method of using the dosecollected in the ACI instead of what was in the capsule to determineFPF.

The terms “FPF (<3.4),” “FPF (<3.4 μm),” and “fine particle fraction ofless than 3.4 m” refers to the fraction of a mass of respirable dryparticles that have an aerodynamic diameter of less than a certain size,e.g., 3.4 μm. For example, FPF (<3.4) can be determined by dividing themass of respirable dry particles deposited on the collection filter of atwo-stage collapsed ACI by the total mass of respirable dry particlesweighed into a capsule for delivery to the instrument. This parametermay also be identified as “FPF_TD (<3.4),” where TD means total dose. Asimilar measurement can be conducted using an eight-stage ACI. Theeight-stage ACI result can also be calculated by the USP method of usingthe dose collected in the ACI instead of what was in the capsule todetermine FPF.

The terms “FPF (<5.0),” “FPF (<5.0 μm),” and “fine particle fraction ofless than 5.0 μm” as used herein, refer to the fraction of a mass ofrespirable dry particles that have an aerodynamic diameter of less thana certain size, e.g., 5.0 μm. For example, FPF (<5.0) can be determinedby using an eight-stage ACI at the standard 60 L/min flow rate byextrapolating from the eight-stage complete data set. This parameter mayalso be identified as “FPF_TD (<5.0),” where TD means total dose.

The portion of the drug formulation falling within a size range istypically referred to as the fine particle fraction (FPF). In somecases, the FPF in the compositions disclosed herein can range from about20% to about 90%, from about 20% to about 80%, from about 20% to about70%, from about 20% to about 60%, from about 20% to about 50%, fromabout 20% to about 40%, from about 20% to about 30%, from about 30% toabout 50%, from about 30% to about 60%, about 30% to about 40%, about37%, or about 44%.

The term “about,” as used herein, refers to a range of +10% of thenumeric value following “about.”

3. Non-Steroidal Anti-Inflammatory Drugs (NSAIDs)

NSAIDs, such as acetylsalicylic acid, can provide various beneficialeffects and contribute to reducing the risk of a cardiovascular disease(such as thrombosis). However, the use of NSAIDs, such asacetylsalicylic acid, in a clinical setting has traditionally beenlimited to oral administration. Oral administration of acetylsalicylicacid, for example, can result in the loss or inactivation ofapproximately ⅔ of the oral dosage due to the first pass effect in thegut and liver. While one third of the dosage reaches the systemic bloodstream and provides the desired effect, the negative side effectscreated by the full dosage often deter patients from usingacetylsalicylic acid on a regular or daily basis.

The methods and systems disclosed herein allow for the beneficialeffects of NSAIDs, such as acetylsalicylic acid, to be achieved on aregular basis and in emergency situations, while minimizing previousdrawbacks associated with the use of NSAIDs.

Various studies have determined that acetylsalicylic acid has asignificant effect on reducing the risk of myocardial infarction. Thesestudies have used acetylsalicylic acid dosages of 325 mg. However, thesestudies have based their findings on oral administration ofacetylsalicylic acid and have not suggested DPI or MDI administration.

Although inhaled dry powder formulations of acetylsalicylic acid havebeen developed, reports have stated that the formulation was notclinically feasible because it is difficult to meet the high dosagerequirements of acetylsalicylic acid (˜80 mg/day for low-dose preventionof coronary events and stroke, and at least 300 mg/day for pain or feverrelief) via pulmonary delivery of dry powders.

In addition, these reports recognize that adverse effects of dry powderon the lungs, such as coughing, cannot be avoided unless the doses areless than a few tenths of a milligram in a single breath. Thus, priorteachings suggest that higher dosage requirements of acetylsalicylicacid would be impossible or difficult to meet using DPI (or MDI).Finally, there may be a higher incidence of acetylsalicylic acidintolerance in asthmatic patients when acetylsalicylic acid is deliveredby inhalation than orally.

The methods and systems disclosed herein provide for treating (includingprophylactic treatment or reducing the risk of) a disease, for example,treating a cardiovascular disease (such as thrombosis) by administrationof a low amount of a NSAID, such as a low dose of acetylsalicylic acid,by DPI. The dose can be much less than that of a baby acetylsalicylicacid (e.g., less than 81 mg). The administered dosage can be less thanabout 40 mg of acetylsalicylic acid. The administered dosage can be lessthan 25 mg of acetylsalicylic acid. Further, the administered dosage canbe less than 20 mg of acetylsalicylic acid. The administered dosage canbe less than 15 mg of acetylsalicylic acid. The administered dosage canalso be less than 12 mg of acetylsalicylic acid. The administered dosagecan be less than 10 mg of acetylsalicylic acid. Furthermore, theadministered dosage can be less than 8 mg of acetylsalicylic acid. Theadministered dosage can be less than 5 mg of acetylsalicylic acid. Insome embodiments, the administered dosage can be less than 2 mg ofacetylsalicylic acid.

For example, the dosage can be from about 1 mg to about 40 mg. Invarious embodiments, the dosage can be from about 4 mg to about 25 mg ofacetylsalicylic acid, about 6 mg to about 20 mg of acetylsalicylic acid,about 8 mg to about 15 mg of acetylsalicylic acid, about 10 mg to about13 mg of acetylsalicylic acid or about 1 mg, about 2 mg, about 3 mg,about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg,about 10 mg, about 11 mg, about 12 mg, about 13 mg, about 14 mg, about15 mg, about 16 mg, about 17 mg, about 18 mg, about 19 mg, or about 20mg of acetylsalicylic acid. Alternatively, the dose of acetylsalicylicacid can be less than about 80 mg, about 1 mg to about 75 mg, about 2 mgto about 60 mg, about 5 mg to about 40 mg, about 10 mg to about 30 mg,about 12 mg to about 25 mg, about 15 mg to about 20 mg, about 60 mg toabout 95 mg, about 50 mg to about 100 mg, about 50 mg to about 80 mg,about 40 mg to about 80 mg, about 20 mg to about 30 mg, about 30 mg toabout 40 mg, about 40 mg to about 50 mg, about 50 mg to about 60 mg,about 60 mg to about 70 mg, about 70 mg to about 80 mg, about 80 mg toabout 90 mg, or about 90 mg to about 100 mg.

In certain embodiments, NSAIDs can be used in various methods andsystems. In some embodiments, NSAIDs can include salicylates, i.e., thesalts and esters of salicylic acid, which have anti-platelet action.Further, NSAIDs can also include one or more of the following compoundslisted in Table 1.

TABLE 1 Examples of NSAIDs Aspirin or acetylsalicylic acid Celecoxib(Celebrex) Dexdetoprofen (Keral) Diclofenac (Voltaren, Cataflam,Voltaren-XR) Diflunisal (Dolobid) Etodolac (Lodine, Lodine XL)Etoricoxib (Algix) Fenoprofen (Fenopron, Nalfron) Firocoxib (Equioxx,Previcox) Flurbiprofen (Urbifen, Ansaid, Flurwood, Proben) Ibuprofen(Advil, Brufen, Motrin, Nurofen, Medipren, Nuprin) Indomethacin(Indocin, Indocin SR, Indocin IV) Ketoprofen (Actron, Orudis, Oruvail,Ketoflam) Ketorolac (Toradol, Sprix, Toradol IV/IM, Toradol IM)Licofelone (under development) Lomoxicam (Xefo) Loxoprofen (Loxonin,Loxomac, Oxeno) Lumiracoxib (Prexige) Meclofenamic acid (Meclomen)Mefenamic acid (Ponstel) Meloxicam (Movalis, Mel ox, Recoxa, Mobic)Nabumetone (Relafen) Naproxen (Aleve, Anaprox, Midol Extended Relief,Naprosyn, Naprelan) Nimesulide (Sulide, Nimalox, Mesulid) Oxaporozin(Daypro, Dayrun, Duraprox) Parecoxib (Dynastat) Piroxicam (Feldene)Rofecoxib (Vioxx, Ceoxx, Ceeoxx) Salsalate (Mono-Gesic, Salflex,Disalcid, Salsitab) Sulindac (Clinoril) Tenoxicam (Mobi flex) Tolfenamicacid (Clotam Rapid, Tufnil) Valdecoxib (B extra)

Other alternatives can also be used instead of a NSAID. Suchalternatives include Plavix (clopidogrel), COX-2 inhibitors, otherremedies such as Nattokinase (an enzyme (EC 3.4.21.62, extracted andpurified from a Japanese food called natto)). Further, other drugs thatprovide different beneficial effects, such as being effective to reducea risk of a cardiovascular disease (such as thrombosis) in a patient,can also be used in some embodiments. Thus, the discussion of methodsand systems shall apply generally to these various alternatives,although for discussion purposes, the present disclosure often refers toacetylsalicylic acid. It is contemplated that the methods, effects,pharmacokinetic data, and other considerations relating toacetylsalicylic acid can be equally applied to other NSAIDs.

4. Dry Powders and Dry Particles

The dry particles of the subject technology may be dispersible. The sizeof the dry particles can be expressed in a variety of ways that areconventional in the art, such as, fine particle fraction (FPF),volumetric median geometric diameter (VMGD), or mass median aerodynamicdiameter (MMAD).

The dry particles of the subject technology may have a VMGD as measuredby HELOS/RODOS at 1.0 bar of about 10 μm or less (e.g., about 0.1 μm toabout 10 μm). Preferably, the dry particles of the subject technologyhave a VMGD of about 9 μm or less (e.g., about 0.1 μm to about 9 μm),about 8 μm or less (e.g., about 0.1 μm to about 8 μm), about 7 μm orless (e.g., about 0.1 μm to about 7 μm), about 6 μm or less (e.g., about0.1 μm to about 6 μm), about 5 μm or less (e.g., less than 5 μm, about0.1 μm to about 5 μm), about 4 μm or less (e.g., 0.1 μm to about 4 μm),about 3 μm or less (e.g., 0.1 μm to about 3 μm), about 2 μm or less(e.g., 0.1 μm to about 2 μm), about 1 μm or less (e.g., 0.1 μm to about1 μm), about 0.5 μm to about 6 μm, about 0.5 μm to about 5 μm, about 0.5μm to about 4 μm, about 0.5 μm to about 3 μm, or about 0.5 μm to about 2μm as measured by HELOS/RODOS at 1.0 bar. In an exemplary embodiment,the dry particles of the subject technology have a VMGD as measured byHELOS/RODOS at 1.0 bar of about 1.3 to about 1.7 μm. In anotherexemplary embodiment, the dry particles of the subject technology have aVMGD as measured by HELOS/RODOS at 1.0 bar of about 0.5 μm to about 2μm.

Alternatively, the dry particles may have a VMGD as measured byHELOS/RODOS at 1.0 bar of about 30 μm or less (e.g., about 5 μm to about30 μm). Preferably, the dry particles of the subject technology have aVMGD of about 25 μm or less (e.g., about 5 μm to about 25 μm), about 20μm or less (e.g., about 5 μm to about 20 μm), about 15 μm or less (e.g.,about 5 μm to about 15 μm), about 12 μm or less (e.g., about 5 μm toabout 12 μm), about 10 μm or less (e.g., about 5 μm to about 10 μm), orabout 8 μm or less (e.g., 6 μm to about 8 μm) as measured by HELOS/RODOSat 1.0 bar. The dry powders can comprise a mixture of particles havingdifferent sizes.

The respirable dry particles can have an MMAD of about 10 μm or less,such as an MMAD of about 0.5 μm to about 10 μm, about 1 μm to about 10μm, about 0.5 μm to about 5 μm, or about 1 μm to about 5 μm. Preferably,the dry particles of the subject technology have an MMAD of about 5 μmor less (e.g. about 0.5 μm to about 5 μm, preferably about 1 μm to about5 μm), about 4 μm or less (e.g., about 1 μm to about 4 μm), about 3.8 μmor less (e.g. about 1 μm to about 3.8 μm), about 3.5 μm or less (e.g.about 1 μm to about 3.5 μm), about 3.2 μm or less (e.g. about 1 μm toabout 3.2 μm), about 3 μm or less (e.g. about 1 μm to about 3.0 μm),about 2.8 μm or less (e.g. about 1 μm to about 2.8 μm), about 2.2 μm orless (e.g. about 1 μm to about 2.2 μm), about 2.0 μm or less (e.g. about1 μm to about 2.0 μm) or about 1.8 μm or less (e.g. about 1 micron toabout 1.8 μm).

Alternatively, the dry powders and dry particles of the subjecttechnology have a FPF of less than 5.0 μm (FPF_TD<5.0 μm) of at leastabout 20%, at least about 30%, at least about 45%, at least about 40%,at least about 45%, at least about 50%, at least about 60%, at leastabout 65% or at least about 70%. Alternatively or in addition, the drypowders and dry particles of the subject technology have a FPF of lessthan 5.0 μm of the emitted dose (FPF_ED<5.0 μm) of at least about 45%,at least about 50%, at least about 60%, at least about 65%, at leastabout 70%, at least about 75%, at least about 80%, or at least about85%.

In another embodiment, the dry powders and dry particles of theinvention can have an FPF of less than about 5.6 μm (FPF <5.6 μm) of atleast about 20%, at least about 30%, at least about 40%, preferably atleast about 45%, at least about 50%, at least about 55%, at least about60%, at least about 65%, or at least about 70%.

In a third embodiment, embodiment, the dry powders and dry particles ofthe invention can have an FPF of less than about 3.4 μm (FPF <3.4 μm) ofat least about 20%, preferably at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at leastabout 50%, or at least about 55%.

The dry powders and dry particles may have a tap density of about 0.1g/cm³ to about 1.0 g/cm³. For example, the small and dispersible dryparticles have a tap density of about 0.1 g/cm³ to about 0.9 g/cm³,about 0.2 g/cm³ to about 0.9 g/cm³, about 0.2 g/cm³ to about 0.9 g/cm³,about 0.3 g/cm³ to about 0.9 g/cm³, about 0.4 g/cm³ to about 0.9 g/cm³,about 0.5 g/cm³ to about 0.9 g/cm³, or about 0.5 g/cm³ to about 0.8g/cm³, greater than about 0.4 g/cc, greater than about 0.5 g/cc, greaterthan about 0.6 g/cc, greater than about 0.7 g/cc, about 0.1 g/cm³ toabout 0.8 g/cm³, about 0.1 g/cm³ to about 0.7 g/cm³, about 0.1 g/cm³ toabout 0.6 g/cm³, about 0.1 g/cm³ to about 0.5 g/cm³, about 0.1 g/cm³ toabout 0.4 g/cm³, about 0.1 g/cm³ to about 0.3 g/cm³, less than 0.3g/cm³. In a preferred embodiment, tap density is greater than about 0.4g/cm³; in another preferred embodiment, tap density is greater thanabout 0.5 g/cm³. Alternatively, tap density may be less than about 0.4g/cm³.

The dry powders and dry particles can have a water or solvent content ofless than about 15% by weight of the dry particle. For example, the dryparticles can have a water or solvent content of less than about 15% byweight, less than about 13% by weight, less than about 11.5% by weight,less than about 10% by weight, less than about 9% by weight, less thanabout 8% by weight, less than about 7% by weight, less than about 6% byweight, less than about 5% by weight, less than about 4% by weight, lessthan about 3% by weight, less than about 2% by weight, less than about1% by weight or be anhydrous. In another embodiment, the dry particlesof the subject technology can have a water or solvent content of lessthan about 6% and greater than about 1%, less than about 5.5% andgreater than about 1.5%, less than about 5% and greater than about 2%,about 2%, about 2.5%, about 3%, about 3.5%, about 4%, about 4.5% about5%.

Depending on the specific applications, the present composition (e.g.,the present dry powders or dry particles) may contain a low or highpercentage of active ingredient in the composition. For example, the dryparticles may contain 3% or more, 5% or more, 10% or more, 15% or more,20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 50% ormore, 60% or more, 70% or more, 75% or more, 80% or more, 85% or more,90% or more, 95% or more, about 30% to about 99%, about 40% to about99%, about 50% to about 99%, about 60% to about 99%, about 70% to about99%, about 80% to about 99%, about 40% to about 95%, or about 50% toabout 95% (weight percentage, w/w) of the active ingredient (e.g.,acetylsalicylic acid).

5. Delivery of Dry Powders

In a dry powder inhalation technique, a patient can use a dry powderinhaler to inhale a powder formulation of a drug, such as a NSAID. Thedose is effective to reduce a risk of a thromboembolic event in thepatient.

Various types of inhalers can be used to provide the drug using a DPI orMDI delivery system. The dose administered can be effective to reduce arisk of a thromboembolic event in a patient.

For example, the dry powder inhaler 10 can comprise a mouthpiece, areservoir for receiving the NSAID, and an actuation member for makingavailable the NSAID for inhalation by a patient through the mouthpiece.

The methods and systems disclosed herein may be adapted for use with anyDPI or MDI device, including, but not limited to Aerolizer®, Diskus®,Ellipta®, Flexhaler®, Handihaler®, Neohaler®, Pressair®, Rotahaler®,Turbuhaler® or Twisthaler® Plastiape, CDMHaler (see,http://www.nationaljewish.org/healthinfo/medications/devices/dry-powder).

The methods and systems of the invention provide an apparatus and methodfor providing a therapeutically effective dose of an NSAID in order toreduce the risk of a thromboembolic event. As discussed above, thegeneral approach is to deliver an NSAID in a pharmaceutically acceptablepowdered form (e.g., Acetylsalicylic acid, and/or derivatives thereof)by means of an inhaler.

With respect to the particle size distribution (PSD), the presentcomposition may contain particles having same (or similar) sizedistribution, or particles having different size distributions. Theparticle sizes of the present composition may have a mono-modal, bimodalor multimodal distribution. As a result, the present composition mayproduce mono-modal, bimodal or multimodal absorption. In other words,administration of the present composition may result in a mono-modal,bimodal or multimodal concentration-time profile.

For example, the present composition may contain one, two or threegroups of the following: particles with a median aerodynamic diameter ina range from about 1 μm to about 5 μm, particles with a medianaerodynamic diameter in a range from about 5 μm to about 15 μm, andparticles with a median aerodynamic diameter greater than about 15 μm,

Mixing particles of the same active ingredient (e.g., acetylsalicylicacid), using batches of particles having different size distributions,may reduce bridging. For example, while a composition having arelatively uniform particle size will aggregate, providing a blendedcomposition having some particles with a median aerodynamic diameter ina range from about 1 μm to about 5 μm, other particles with a medianaerodynamic diameter in a range from about 5 μm to about 15 μm, andstill other particles with a median aerodynamic diameter greater thanabout 15 μm, may inhibit aggregation and maintain the depositioncharacteristics of the preparation. In effect, the pharmaceuticallyactive compound is used to replace the function of an excipient (such aslactose) with respect to preventing aggregation during storage of themedicament.

In addition, by selecting the proportions of the various particle sizes,one can provide formulations that are faster or slower acting, based onthe location of where the drug is ultimately deposited. For example,some embodiments provide a preparation that comprises 80%acetylsalicylic acid particles with a median aerodynamic diameter ofabout 1 μm to about 5 μm, and about 20% of particles with a medianaerodynamic diameter of at least 15 μm. Other combinations are possibleas well, and those of skill in the art will readily appreciate thatfaster acting preparations will comprise proportionately more smallerparticles, while slower acting preparations will compriseproportionately more large particles. Thus, using the apparatus andmethods described herein it is therefore possible to provide atherapeutically effective dose of an NSAID such as acetylsalicylic acidvia the respiratory tract, at least as rapidly as can be achieved byoral dosing.

Where a slower acting dosage form is desired, the formulation mayinclude increasing fractions of particles with a median aerodynamicdiameter in the range from about 5 μm to about 10 μm, or 15 μm orgreater. These preparations would result in deposition in either theairways or oral cavity and pharynx and thus provide a more gradualincrease in circulating levels of acetylsalicylic acid and its metabolicderivatives.

Accordingly, one aspect of the subject technology provides a dry powderthat comprises a mixture of particles of various sizes.

For example, the dry powder can comprise particles of large sizes, asmeasured by VMGD (e.g., VMGD≥15 μm, such as ≥20 μm or 20-30 μm) and ofsmall sizes, as measured by VMGD (e.g., VMGD≤5 μm, such as 1-3 μm) at aratio (w:w) of: about 1:1, about 1:2, about 1:3, about 1:4, about 1:5,about 1:6, about 1:7, about 1:8, about 1:10, about 1:15, about 1:20,about 1:25, about 1:30, about 1:40, about 1:50, about 1:100, about 2:1,about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about9:1, about 10:1, about 15:1, about 20:1, about 25:1, about 30:1, about40:1, about 50:1, or about 100:1, etc.

Alternatively, the dry powder can comprise: about 1%, about 5%, about10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%,about 45%, about 50%, about 55%, about 50%, about 55%, about 60%, about65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%,or about 99% (weight percentage) of particles having VMGD of about 10 μmor less, preferably about 5 μm or less. Particles of 10 μm or lessgenerally can reach lungs, and particles of 5 μm or less (e.g., 1-3 μm)generally can reach alveoli.

In another embodiment, the dry powder can comprise: about 1%, about 5%,about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about40%, about 45%, about 50%, about 55%, about 50%, about 55%, about 60%,about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about95%, or about 99% (weight percentage) of particles having VMGD ofbetween about 5 μm to about 20 μm, preferably between about 5 μm toabout 15 μm, or between about 5 μm to about 10 μm.

Alternatively, the dry powder can comprise: about 1%, about 5%, about10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%,about 45%, about 50%, about 55%, about 50%, about 55%, about 60%, about65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%,or about 99% (weight percentage) of particles having VMGD of about 15μm, 20 μm or more.

The above features can be combined. For example, the dry power cancomprise about 50% of particles of about 5 μm or less (VMGD), about 25%of particles of about 5 to about 15 μm (VMGD), and about 25% ofparticles of about 15 μm or more (VMGD).

The dry powder can also comprise a mixture of particles having variousmass median aerodynamic diameters (MMAD). For example, the dry powdercan comprise particles of large sizes (e.g., MMAD 2: 15 μm, such as 2:20μm or 20-30 μm) and of small sizes (e.g., MMAD: 5 μm, such as 1-3 μm) ata ratio (w:w) of: about 1:1, about 1:2, about 1:3, about 1:4, about 1:5,about 1:6, about 1:7, about 1:8, about 1:10, about 1:15, about 1:20,about 1:25, about 1:30, about 1:40, about 1:50, about 1:100, about 2:1,about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about9:1, about 10:1, about 15:1, about 20:1, about 25:1, about 30:1, about40:1, about 50:1, or about 100:1, etc

Alternatively, the dry powder can comprise: about 1%, about 5%, about10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%,about 45%, about 50%, about 55%, about 50%, about 55%, about 60%, about65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%,or about 99% (weight percentage) of particles having MMAD of about 10 μmor less, preferably about 5 μm or less. Particles of 10 μm or lessgenerally can reach lungs, and particles of 5 μm or less (e.g., 1-3 μm)generally can reach alveoli.

In another embodiment, the dry powder can comprise: about 1%, about 5%,about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about40%, about 45%, about 50%, about 55%, about 50%, about 55%, about 60%,about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about95%, or about 99% (weight percentage) of particles having MMAD ofbetween about 5 μm to about 20 μm, preferably between about 5 μm toabout 15 μm, or between about 5 μm to about 10 μm.

In another embodiment, the dry powder can comprise: about 1%, about 5%,about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about40%, about 45%, about 50%, about 55%, about 50%, about 55%, about 60%,about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about95%, or about 99% (weight percentage) of particles having MMAD of about15 μm or more, preferably 20 μm or more.

The above features can be combined. For example, the dry power cancomprise about 50% of particles of about 5 μm or less (MMAD), about 25%of particles of about 5 to about 15 μm (MMAD), and about 25% ofparticles of about 15 μm or more (MMAD).

In certain embodiments, the dry powder may not have an excipient, whichmay be an anti-aggregation (or anti-bridging) excipient.

The dry powder can comprise a mixture of particles of various sizes, andis effective to substantially prevent or reduce particle bridging. Incertain embodiments, at least about 40%, at least about 45%, at leastabout 50%, at least about 55%, at least about 60%, at least about 65%,at least about 70%, at least about 75%, at least 80%, at least about85%, or at least about 90% of the NSAID (such as acetylsalicylic acid)in the dry powder is delivered to the alveolar spaces of a lung.

6. Methods for Preparing Dry Powders and Dry Particles

The dry particles and dry powders can be prepared using any suitablemethod. Many suitable methods for preparing dry powders and particlesare conventional in the art, and include single and double emulsionsolvent evaporation, spray drying, milling (e.g., jet milling),blending, solvent extraction, solvent evaporation, phase separation,simple and complex coacervation, interfacial polymerization, suitablemethods that involve the use of supercritical carbon dioxide (CO₂), andother suitable methods. Dry particles can be made using methods formaking microspheres or microcapsules known in the art. These methods canbe employed under conditions that result in the formation of dryparticles having the desired aerodynamic properties (e.g., aerodynamicdiameter and geometric diameter). If desired, dry particles with desiredproperties, such as size and density, can be selected using suitablemethods, such as sieving.

Spray Drying

Dry particles can be produced by spray drying. Suitable spray dryingtechniques are described, for example, by K. Masters in “Spray DryingHandbook”, John Wiley & Sons, New York (1984); and spray dryingtechniques developed by BUCHI Laboratory Equipment or GEA Niro dryingtechnology. Generally, during spray drying, heat from a hot gas such asheated air or nitrogen is used to evaporate a solvent from dropletsformed by atomizing a continuous liquid feed. If desired, the spraydrying or other instruments, e.g., jet milling instrument, used toprepare the dry particles can include an inline geometric particle sizerthat determines a geometric diameter of the respirable dry particles asthey are being produced, and/or an inline aerodynamic particle sizerthat determines the aerodynamic diameter of the respirable dry particlesas they are being produced.

For spray drying, solutions, emulsions or suspensions that contain thecomponents of the dry particles to be produced in a suitable solvent(e.g., aqueous solvent, organic solvent, aqueous-organic mixture oremulsion) are distributed to a drying vessel via an atomization device.For example, a nozzle or a rotary atomizer may be used to distribute thesolution or suspension to the drying vessel. For example, a rotaryatomizer having a 4- or 24-vaned wheel may be used. Examples of suitablespray dryers that can be outfitted with either a rotary atomizer or anozzle, include, Mobile Minor Spray Dryer or the Model PSD-1, bothmanufactured by Niro, Inc. (Denmark). Actual spray drying conditionswill vary depending, in part, on the composition of the spray dryingsolution or suspension and material flow rates. The person of ordinaryskill will be able to determine appropriate conditions based on thecompositions of the solution, emulsion or suspension to be spray dried,the desired particle properties and other factors. In general, the inlettemperature to the spray dryer is about 100° C. to about 300° C., andpreferably is about 220° C. to about 285° C. The spray dryer outlettemperature will vary depending upon such factors as the feedtemperature and the properties of the materials being dried. Generally,the outlet temperature is about 50° C. to about 150° C., preferablyabout 90° C. to about 120° C., or about 98° C. to about 108° C. Ifdesired, the respirable dry particles that are produced can befractionated by volumetric size, for example, using a sieve, orfractioned by aerodynamic size, for example, using a cyclone, and/orfurther separated according to density using techniques known to thoseof skill in the art.

To prepare the dry particles of the subject technology, generally, asolution, emulsion or suspension that contains the desired components ofthe dry powder (i.e., a feed stock) is prepared and spray dried undersuitable conditions. Preferably, the dissolved or suspended solidsconcentration in the feed stock is at least about 1 g/L, at least about2 g/L, at least about 5 g/L, at least about 10 g/L, at least about 15g/L, at least about 20 g/L, at least about 30 g/L, at least about 40g/L, at least about 50 g/L, at least about 60 g/L, at least about 70g/L, at least about 80 g/L, at least about 90 g/L, or at least about 100g/L. The feedstock can be provided by preparing a single solution orsuspension by dissolving or suspending suitable components (e.g., salts,excipients, other active ingredients) in a suitable solvent. Thesolvent, emulsion or suspension can be prepared using any suitablemethods, such as bulk mixing of dry and/or liquid components or staticmixing of liquid components to form a combination. For example, ahydrophilic component (e.g., an aqueous solution) and a hydrophobiccomponent (e.g., an organic solution) can be combined using a staticmixer to form a combination. The combination can then be atomized toproduce droplets, which are dried to form respirable dry particles.Preferably, the atomizing step is performed immediately after thecomponents are combined in the static mixer.

In one example, dry particles that comprise acetylsalicylic acid can beprepared by spray drying. Spray drying is a commonly used method ofdrying a liquid feed through a hot gas. It is a method whereby solutionsor slurries can be rapidly dried to particulate form by atomizing theliquid in a heated chamber. Typically, the hot gas can be air. Whenpreparing chemically sensitive materials such as pharmaceuticals, orwhen solvents such as ethanol are used, oxygen-free atmosphere may berequired, such as nitrogen. Spray drying is frequently used in the foodpreparation industry and has become an important method for thedehydration of fluid foods such as milk, coffee, and egg powder. Theprocess is also adaptable to preparations of pharmaceutical and chemicalformulations.

The liquid feed varies depending on the material being dried and is notlimited to food or pharmaceutical products, and may be a solution,colloid or suspension. The process is a one-step rapid method thattypically eliminates additional processing. By controlling processconditions particles of the desired size can be reproducibly formed.

The spray drying can be conducted in the presence or absence of one ormore excipients.

In some cases, excipients can be included with the active pharmaceuticalingredient such that a complex particle of active pharmaceuticalingredient (API) and excipient can be produced in a single step process.In other cases, an active pharmaceutical particulate preparation can beproduced in a first spray-drying process, and that product then modifiedby the subsequent addition of one or more pharmaceutically acceptableexcipients. In some cases, it is possible to add excipients by asubsequent spray-drying process.

In some spray-drying methods, the liquid feed is pumped through anatomizer nozzle, or array of nozzles, that produce fine droplets thatare introduced into the main drying chamber. Atomizers can vary therebeing rotary, single fluid, two-fluid, and ultrasonic designs. Thesedifferent designs provide a variety of advantages, applicability anddisadvantages depending on the particular spray drying process required.The hot drying gas can be passed as a concurrent or counter-current flowto the atomizer direction. The concurrent flow enables the particles tohave a lower residence time within the system and the particle separatorthus operates more efficiently. In some systems the particle separatoris a cyclone device. The counter-current flow method enables a greaterresidence time of the particles in the chamber. Therefore, in general aspray-drying method will consist of the steps of pre-concentration ofliquid, atomization, drying in a hot gas atmosphere, separation of thedried powder from moist gas, cooling, and then packaging of the finishedproduct.

In one embodiment, feed solutions with acetylsalicylic acidconcentrations of either 2% w/w, or 5% w/w, were prepared by addingacetylsalicylic acid to the appropriate solvent followed by stirringuntil a homogeneous solution was obtained. A BUCHI spray dryer modelB-290 Advanced was used in all experiments. The unit was equipped with atwo fluid nozzle where the nozzle and diameter were 1.4 mm and 0.7 mm,respectively. The high-performance cyclones were used to collect thedried product. The spray-drying unit was operated in open cycle, withthe aspirator blowing nitrogen at 100% of capacity, corresponding to aflow rate of the dry nitrogen of approximately 40 kg per hour. The flowrate of atomization nitrogen was adjusted to 40 mm or 50 mm in therotameter, depending on the particular trial. Before feeding the stocksolution, the spray dryer was stabilized with the solvent. During thestabilization period, the solvent flow rate was adjusted in order togive the target outlet temperature. After stabilization of the outlettemperature, the feed of the spray dryer was commuted from the solventto the product solution (inlet temperature was then readjusted tomaintain the outlet temperature in the target value). At the end of thestock solution, the feed was once more commuted to solvent, in order torinse the feed line and carry out a controlled shutdown.

The initial objective of these experiments was to isolate the amorphousform of acetylsalicylic acid, in order to fully characterize it.However, as was discovered from a review of the literature,acetylsalicylic acid presents a negative Tg (of −30° C.), and as suchthe option of producing a crystalline size reduced active pharmaceuticalwith this technique was attempted. For that purpose, for solutions ofacetylsalicylic acid in ethanol (the most suited solvent to dissolve theacetylsalicylic acid, given its high solubility and its approval forinhalation use) were prepared and spray dried as follows. Inlettemperature ranged from about 80° C. to about 160° C. Outlet temperaturewas initially set to 65° C. In one experiment the outlet temperature wasincreased to 100° C. in an attempt to accelerate theamorphous-crystalline conversion, in the hopes that this would reducelosses that are typical of the transient glassy state of the material.However, increasing the outlet temperature did not produce anyappreciable increase in overall yield of product. The rotameter wasvaried from about 40 mm to about 50 mm. Feed rate was typically about 5mL per minute. Following spray drying, a number of analytical methodswere used to evaluate the resulting product.

X-ray powder diffraction (XRPD) showed that in each of the fourdifferent batches prepared acetylsalicylic acid appeared to becrystalline in form, and the diffractogram was similar to that of thestarting material. In addition, the spray dried products presentedthermal grams that were identical to the input material. The overallyield ranged from about 55% to about 65%.

The melting temperature of the resulting spray dried product ranged fromabout 133° C. to about 137° C., comparing favorably with the publishedmelting point for acetylsalicylic acid (136° C.). A measure ofhygroscopic properties showed a weight change ranging from −0.4% toabout 1.2% when the products were exposed to an atmosphere with 95%relative humidity. These results suggest no issues with hygroscopicbehavior and that with respect to this property, spray driedacetylsalicylic acid behaves in a manner similar to that of unprocessedacetylsalicylic acid.

Particle size distribution analysis showed that DV₁₀ ranged from about0.9 μm to about 1.2 μm, DV₅₀ ranged from about 3 μm to about 6 μm, andDV90 ranged from about 8 μm to about 24 μm. It was discovered that byreducing feed concentration of acetylsalicylic acid to 2% w/w, a smalleraverage particle size could be obtained, which was within typicalinhalation range.

HPLC analysis showed acetylsalicylic acid purity to range from about 92%to about 98%, with the major “impurity” being salicylic acid, whichranged from about 0.3% to about 0.5%. Residual solvent ranged from about90 ppm to about 150 ppm, well below the limits defined in the ICH Q3Aguidelines.

The feedstock, or components of the feed-stock, can be prepared usingany suitable solvent, such as an organic solvent, an aqueous solvent ormixtures thereof. Suitable organic solvents that can be employed includebut are not limited to alcohols such as, for example, ethanol, methanol,propanol, isopropanol, butanols, and others. Other organic solventsinclude but are not limited to perfluorocarbons, dichloromethane,chloroform, ether, ethyl acetate, methyl tert-butyl ether and others.Co-solvents that can be employed include an aqueous solvent and anorganic solvent, such as, but not limited to, the organic solvents asdescribed above. Aqueous solvents include water and buffered solutions(such as phosphate buffer).

The feedstock or components of the feed stock can have any desired pH,viscosity or other properties. If desired, a pH buffer can be added tothe solvent or co-solvent or to the formed mixture. Generally, the pH ofthe mixture ranges from about 3 to about 8.

Jet Milling

Respirable particles can also be produced by jet-milling. See, e.g.,techniques developed by Apex Process Technology or Jetpharma SA. Jetmilling is a process of using highly compressed air or other gasses,usually in a vortex motion, to impact fine particles against each otherin a chamber. Jet mills are capable of reducing solids to particle sizesin the low-micron to submicron range. The grinding energy is created bygas streams from horizontal grinding air nozzles. Particles in thefluidized bed created by the gas streams are accelerated towards thecenter of the mill, colliding with slower moving particles. The gasstreams and the particles carried in them create a violent turbulenceand as the particles collide with one another they are pulverized.

In certain embodiments, jet-milling was able to produce acetylsalicylicacid particles with a FPF within the desired inhalable range for maximaldeposition at the deepest levels of the lung. In some cases the DV90 wasless than about 9 μm, in some cases less than about 5 μm, and in somecases less than about 3 μm. Particles produced by jet milling can beefficiently and predictably delivered from a dry powder inhaler device,and at least 25% of the particles are of a size that would be expectedto deposit within the alveolar spaces of the lungs. In some cases atleast 50% of the particles are of a size that would be expected todeposit within the alveolar spaces of the lungs. In one embodiment, atleast 75% of the particles are of a size that would be expected todeposit within the alveolar spaces of the lungs. In another embodiment,at least 90% of the particles are of a size that would be expected todeposit within the alveolar spaces of the lungs.

Wet Polishing

Wet polishing is a process that combines a technology to attain a smallparticle size (either a bottom up technique such as controlledcrystallization or nanocrystallization or top down technique such ashigh shear mixing or high pressure homogenization) with a suitableisolation technology (for example spray drying or filtration with adrying process). See, e.g., techniques developed by Hovione. Thesecombinations can be used to tune the particle size and morphology tomeet specific drug delivery needs. The method allows control of particlesize distribution with tight spans and in-process sampling, andmaintains crystalline state (little or no amorphous content).

Wet polishing technique can be repeated multiple times to achieve aparticular size of about 500 nanometers or less. Studies were undertakento investigate whether wet polishing could provide an appropriate methodfor producing acetylsalicylic acid particles of an inhalable size andwhich were deliverable from a dry powder inhaler device. Initially, aliterature review was conducted in order to determine the best candidateanti-solvent for use in reducing acetylsalicylic acid particle size by awet-polishing method. Solvents were evaluated by their predicted abilitywith respect to minimal solubility of acetylsalicylic acid. From thisreview, the following candidate solvents were identified: water,benzene, toluene, hexane, n-heptane, dibutyl ether anddi-isopropyl-ether.

After considering a number of factors in the end it was determined thatonly n-heptane and toluene fulfilled all the requirements, and thereforethese were selected for further evaluation. Suspensions ofacetylsalicylic acid at 5% w/w were prepared with the variousanti-solvents, by charging the required amount of anti-solvent, chargingthe required amount of acetylsalicylic acid, and then stirring until ahomogeneous suspension was obtained. The suspensions were qualitativelyevaluated at room temperature and then filtered using a 0.45 μmmembrane, which was then placed in an oven and dried at 60° C. until thesolvent had completely evaporated. A quantitative analysis was performedon the residue that remained in the membrane by weighing the membranebefore and after the test. From this analysis it was determined thatacetylsalicylic acid was partially soluble in toluene, and displayed aphobic behavior towards n-heptane.

Suspensions were prepared using either toluene or n-heptane at anacetylsalicylic acid concentration of 5% w/w. Each individual suspensionwas then subjected to a milling operation using a Microfluidics ModelM-110EH-30 apparatus. Milling was conducted at a pressure of 50 barusing a 200 μm chamber for between 20 and 70 cycles. Input temperaturesranged from about 80° C. to about 140° C. Output temperatures rangedfrom about 65° C. to about 90° C. The process yield ranged from about 5%to about 25%.

Analysis of the resulting product revealed a DV₁₀ ranging from about 1.5μm to about 3.3 μm, DV₅₀ ranging from about 3.3 μm to about 6.7 μm, andDV90 ranging from about 6.3 μm to about 12.0 μm. HPLC assay of the finalproduct revealed a composition of acetylsalicylic acid ranging from 90%to about 98%, with impurities ranging from about 1.4% to about 12%. Theprimary impurity was salicylic acid. It was also observed that all ofthe products obtained by wet polishing were more hygroscopic than theraw material, showing a water gain of about 5% when in the presence of95% relative humidity.

In addition, when tested for aerodynamic performance, acetylsalicylicacid processed by wet polishing performed poorly in two different drypowder delivery devices. When examining devices loaded with either 15 mgor 40 mg, a significant amount of material (about 25 to 30%) wasretained within the inhalation device itself. Overall, the resultssuggested that wet polishing alone impacts to a significant extent thephysical and chemical properties of acetylsalicylic acid, and thereforewere seen to be less desirable for producing a pharmaceutical productfor inhalation. Wet polishing may nonetheless be a suitable method formicronization of acetylsalicylic acid for other purposes.

In some cases, it is possible to produce acetylsalicylic acid particlesof a desired size using a process known as controlled crystallization.It is well known in the art that the crystalline state of most compoundsis more thermodynamically stable that the amorphous state. As a result,producing acetylsalicylic acid in crystalline form is expected toimprove stability of the active ingredient. In addition, production ofacetylsalicylic acid in crystalline form also provides the potential tomodify the active ingredient to optimize various biochemical properties,such as solubility, dissolution rate and pH solubility profile (amongothers) in order to improve pharmacokinetic performance. In some cases,sequential crystallization steps can be used to improve the purity ofthe active ingredient and selectively remove undesirable impurities.

Similarly, through the proper selection of various solvents andanti-solvents, it is possible to manipulate physical characteristicssuch as crystal shape. It is well known that certain crystal shapes aredifficult to handle both at the product development and manufacturingstages. For example, needles and flakes are widely regards as lessdesirable particle shapes. It is possible however, to manipulate crystalformation in order to direct the final product to more suitable crystalshapes. In some cases, it is possible to grow crystals with high aspectratios using non-polar hydrocarbon solvents such as hexane or heptane.In contrast, crystals having a low aspect ratio can be produced usingpolar solvents such as methanol or ethanol. The addition of surfaceactive “impurities” can also be used to inhibit crystal growth incertain planar forms.

The solubility of acetylsalicylic acid in a number of solvents was firstevaluated prior to initiating controlled crystallization by addition ofa suitable anti-solvent. The results are shown in the following Table 2.

TABLE 2 Solubility of acetylsalicylic acid Solvent T (°C.) g/ml T (°C.)g/ml EtOH 23 0.125 3 0.063 Acetone 23 0.200 3 0.143 MeOH 23 0.167 30.133 DMF 23 0.500 — — THF 23 0.500 3 0.250 PEG-200 23 0.077 — —

Next, several small crystallization experiments were carried out toevaluate the behavior of the acetylsalicylic acid in different systems.Each experiment consisted of dissolving 2 gm of acetylsalicylic acid ina solvent (T=20-25° C.), and then adding this solution to theanti-solvent (100 vol. of anti-solvent at −5° C.). The suspensionobtained was stirred for 15 min and solid material collected byfiltration and then dried. Table 3 summarizes the conditions of eachexperiment.

TABLE 3 Summary of the crystallization experiments Anti- Solvent (v/w) T(° C.) Solvent (v/w) T (° C.) Crystals Yield EtOH 8 20-25 H₂O 100 4 yes51.5 EtOH 8 20-25 n-Hept 100 4 yes 58.0 EtOH 8 20-25 Toluene 100 4 no —EtOH 8 20-25 H₂O 100 4 yes 53.0 H2SO4 0.05 THF 2.5 20-25 H₂O 100 4 yes53.5 THF 2.5 20-25 Toluene 100 4 yes 45.5 THF 2.5 20-25 n-Hept 100 4 yes89.0 MeOH 6 20-25 H₂O 100 4 yes 64.5 MeOH 6 20-25 n-Hept 100 4 yes 17.0MeOH 6 20-25 Toluene 100 4 no — Acetone 7 20-25 H₂O 100 3 yes 43.0Acetone 7 20-25 n-Hept 100 4 yes 71.5 Acetone 7 20-25 Toluene 100 4 yes34.5Excipients

Particles described herein can be encapsulated, e.g., by apharmaceutical excipient such as lactose, sugar, or a polymer.

In addition, particles described herein can be mixed and/or coated withvarious pharmaceutically acceptable excipients. Excipients can beincluded in order to improve aerodynamic performance of the active drug,to improve bioavailability, increase stability, to modulate pH, toprovide sustained release properties, to provide taste-masking of anirritating drug and/or to improve pharmacokinetic performance.

With dry powder formulations, excipients can also provide a carrierfunction to reduce clumping of the active pharmaceutical ingredient andto improve suspension of the formulation in the airflow as thepharmaceutical preparation is being inhaled. Such carriers can includesubstances such as, but not limited to, sugars/sugar alcohols such asglucose, saccharose, lactose and fructose, starches or starchderivatives, oligosaccharides such as dextrins, cyclodextrins and theirderivatives, polyvinylpyrrolidine, alginic acid, tylose, silicic acid,cellulose, cellulose derivatives, sugar alcohols such as mannitol orsorbitol, calcium carbonate, calcium phosphate, lactose, lactitol,dextrates, dextrose, maltodextrin, saccharides includingmonosaccharides, disaccharides, polysaccharides; sugar alcohols such asarabinose, ribose, mannose, sucrose, trehelose, maltose and dextran.

In some cases, an excipient can be provided in order to coat the activepharmaceutical ingredient, thus “masking” it. Masking is especiallyuseful when the unmodified active pharmaceutical is irritating orotherwise unpleasant to the recipient. For example, in some cases it hasbeen shown that coating a bitter molecule with a hydrogenated oil andsurfactant combination is effective to cover the otherwise unpleasanttaste of the active ingredient.

Non-limiting examples of pharmaceutically acceptable excipients includephospholipids, amino acids, polypeptides and combinations thereof. Thephospholipids may or may not have surfactant properties. Examples ofsuitable phospholipid excipients include, without limitation,phosphatidylcholines, phosphatidylethanolamines, phosphatidylinositol,phosphatidylserines, sphingomyelin or other ceramides, as well asphospholipid containing oils such as lecithin oils. Combinations ofphospholipids, or mixtures of a phospholipid(s) and other substance(s),may be used. In one embodiment, the phospholipids used as excipients aresoy lecithin. In another embodiment, the phospholipid is endogenous tothe lung.

Non-limiting examples of the phospholipids that may be used in thepresent composition include, soy lecithin,dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine(DSPC), dilaurylolyphosphatidylcholine (DLPC),dimyristoylphosphatidylcholine (DMPC), dioleoylphosphatidylcholine(DOPC), dilaurylolylphosphatidylglycerol (DLPG),dimyristoylphosphatidylglycerol (DMPG), dipalmitoylphosphatidylglycerol(DPPG), distearoylphosphatidylglycerol (DSPG),dioleoylphosphatidylglycerol (DOPG), dimyristoyl phosphatidic acid(DMPA), dimyristoyl phosphatidic acid (DMPA), dipalmitoyl phosphatidicacid (DPPA), dipalmitoyl phosphatidic acid (DPPA), dimyristoylphosphatidylethanolamine (DMPE), dipalmitoyl phosphatidylethanolamine(DPPE), dimyristoyl phosphatidylserine (DMPS), dipalmitoylphosphatidylserine (DPPS), dipalmitoyl sphingomyelin (DPSP), anddistearoyl sphingomyelin (DSSP).

In one embodiment, soy lecithin, dipalmitoyl phosphatidylcholine (DPPC),distearoyl phosphatidylcholine (DSPC) or a mixture thereof are used asan excipient.

The amino acid and/or polypeptide may be water-soluble. The amino acidmay be an L-amino acid or a D-amino acid. The amino acid may be Leucine,Alanine, Arginine, Asparagine, Aspartic acid, Cysteine, Glutamic acid,Glutamine, Glycine, Histidine, Isoleucine, Lysine, Methionine,Phenylalanine, Proline, Serine, Threonine, Tryptophan, Tyrosine, Valine,or combinations or variants thereof.

An excipient may be used to in the present composition. The excipient(s)may be present at levels ranging from about 0% to about 99% (w/w), fromabout 0.01% to about 80% (w/w), from about 0.05% to about 70% (w/w),from about 0.1% to about 60% (w/w), from about 0.1% to about 50% (w/w),from about 0.1% to about 40% (w/w), from about 0.1% to about 30% (w/w),from about 0.1% to about 20% (w/w), from about 0.1% to about 10% (w/w),from about 0.05% to about 8% (w/w), from about 0.1% to about 6% (w/w),from about 5% to about 10% (w/w), from about 3% to about 8% (w/w), fromabout 2% to about 6% (w/w), from about 0.1% to about 5% (w/w), fromabout 0.1% to about 4% (w/w), from about 0.1% to about 3% (w/w), fromabout 0.1% to about 2% (w/w), from about 0.1% to about 1% (w/w), fromabout 1% to about 6% (w/w), from about 1% to about 5% (w/w), from about1% to about 4% (w/w), or from about 1% to about 3% (w/w) of theparticles. In certain embodiments, one or more excipients (e.g., one ormore phospholipids, amino acids, and/or polypeptides) are present atlevels in a range from about 0.1% (w/w) to about 10% (w/w), from about0.1% (w/w) to about 40% (w/w), from about 1% to about 30% (w/w), fromabout 0.5% to about 20% (w/w), from about 0% to about 99% (w/w), fromabout 0.01% to about 80% (w/w), from about 0.05% to about 70% (w/w),from about 0.1% to about 60% (w/w), from about 0.1% to about 50% (w/w),from about 0.1% to about 40% (w/w), from about 0.1% to about 30% (w/w),from about 0.1% to about 20% (w/w), from about 0.05% to about 8% (w/w),from about 0.1% to about 6% (w/w), from about 5% to about 10% (w/w),from about 3% to about 8% (w/w), from about 2% to about 6% (w/w), fromabout 0.1% to about 5% (w/w), from about 0.1% to about 4% (w/w), fromabout 0.1% to about 3% (w/w), from about 0.1% to about 2% (w/w), fromabout 0.1% to about 1% (w/w), from about 1% to about 6% (w/w), fromabout 1% to about 5% (w/w), from about 1% to about 4% (w/w), from about1% to about 3% (w/w), about 0.1%, about 5% (w/w), about 4% (w/w), about3%, about 10% (w/w), about 13% (w/w), or about 15% (w/w), of theparticles.

In addition, in some embodiments, the surfactant can be provided incombination with one or more additional excipients including absorbents,acidifiers, alkalizers, buffers, antimicrobial agents, antioxidants,binders, solubilizing agents, solvents, viscosity modifiers, humectantsand combinations thereof. In some embodiments the formulation includessalts in amounts effective to render the dissolved formulation isosmoticwith the lung.

Respirable dry particles and dry powders can be fabricated and thenseparated, for example, by filtration or centrifugation by means of acyclone, to provide a particle sample with a preselected sizedistribution. For example, greater than about 30%, greater than about40%, greater than about 50%, greater than about 60%, greater than about70%, greater than about 80%, or greater than about 90% of the respirabledry particles in a sample can have a diameter within a selected range.The selected range within which a certain percentage of the respirabledry particles fall can be, for example, any of the size ranges describedherein, such as between about 0.1 to about 3 μm VMGD.

The diameter of the respirable dry particles, for example, their VMGD,can be measured using an electrical zone sensing instrument such as aMultisizer lie, (Coulter Electronic, Luton, Beds, England), or a laserdiffraction instrument such as a HELOS system (Sympatec, Princeton,N.J.). Other instruments for measuring particle geometric diameter arewell known in the art. The diameter of respirable dry particles in asample will range depending upon factors such as particle compositionand methods of synthesis. The distribution of size of respirable dryparticles in a sample can be selected to permit optimal depositionwithin targeted sites within the respiratory system.

Experimentally, aerodynamic diameter can be determined using time offlight (TOF) measurements. For example, an instrument such as the Model3225 Aerosizer DSP Particle Size Analyzer (Amherst Process Instrument,Inc., Amherst, Mass.) can be used to measure aerodynamic diameter. TheAerosizer measures the time taken for individual respirable dryparticles to pass between two fixed laser beams.

Aerodynamic diameter also can be experimentally determined directlyusing conventional gravitational settling methods, in which the timerequired for a sample of respirable dry particles to settle a certaindistance is measured. Indirect methods for measuring the mass medianaerodynamic diameter include the Andersen Cascade Impactor (ACI) and themulti-stage liquid impinger (MSLI) methods. Another method of measuringthe aerodynamic diameter is with a Next Generation Impactor (NGI). TheNGI operates on similar principles of inertial impaction as the ACI. TheNGI consists of seven stages and can be calibrated at flow rates of 30,60, and 100 LPM. In contrast to the ACI, for which the impactor stagesare stacked, the stages of the NGI are all in one plane. Collection cupsare used to collect the particles below each stage of the NGI. U.S. Pat.No. 8,614,255. The methods and instruments for measuring particleaerodynamic diameter are well known in the art.

The dry powder of the present formulation comprises substantially dryparticles having a mass median aerodynamic diameter (MMAD) within arange of about 0.5 μm to about 10 μm, wherein the dry powder furthercomprises one or more phospholipids in an amount ranging from about 0.1%(w/w) to about 10% (w/w) of the dry particles. The particles may have anMMAD size distribution where the particles exhibit: (i) a DV90 less thanabout 20 μm, a DV50 less than about 7 μm, and a DV10 less than about 2μm; (ii) a DV90 less than about 10 μm, a DV50 less than about 4 μm, anda DV10 less than about 1 μm; or (iii) a DV90 less than about 6 μm, aDV50 less than about 3 μm, and a DV10 less than about 1 μm.

The dry powder may comprise particles coated with a pharmaceuticallyacceptable excipient. The pharmaceutically acceptable excipient can be aphospholipid having surfactant properties, such as dipalmitoylphosphatidylcholine (DPPC), distearoyl phosphatidylcholine (DSPC) or soylecithin, in an amount ranging from about 0.1% to about 10% w/w or in anamount ranging from about 1% to about 5% w/w. In one embodiment, theweight percentage of the DSPC on the dry particles is 5% w/w. In anotherembodiment, the respirable dry powder of the soy lecithin of the dryparticles is 0.1% w/w.

The drug delivery system disclosed herein is effective in reducing therisk of a thromboembolic event or treat thrombosis. The dose ofacetylsalicylic acid may be present at amounts ranging from about 5 toabout 40 mg. The formulation my further comprise clopidogrel. Therespirable dry powders can have an emitted dose ranging from about 75%to about 95%.

In one embodiment, the pharmaceutically acceptable excipient is DSPC,the respirable dry powders substantially comprise dry particles having aMMAD ranging from about 3 to about 4 μm and an emitted dose greater thanabout 90%. In this embodiment, the mass percent of stages in an NGItesting apparatus of the respirable powder yields are at, stage 1 about10% to about 13%, stage 2, about 20% to about 23%, stage 3, about 13% toabout 15%, and stage 4, about 5% to about 6% and fine particle fractionranges from about 45% to about 55%. Ranges between 80%-120% of actualpercentages as set forth above are encompassed within each embodiment.

In another embodiment, the pharmaceutically acceptable excipient is soylecithin, the respirable dry powders substantially comprise dryparticles having a MMAD ranging from about 2.0 to about 3.0 μm and anemitted dose ranging from about 70% to about 85%. In this embodiment,the mass percent of stages in an NGI testing apparatus of the respirablepowder yields are at, stage 1 about 5% to about 10%, stage 2, about 10%to about 18%, stage 3, about 15% to about 20%, and stage 4, about 10% toabout 15% and fine particle fraction ranges from about 50% to about 70%.Ranges between 80%-120% of actual percentages set forth above areencompassed within each embodiment.

Tap density is a measure of the envelope mass density characterizing aparticle. The envelope mass density of a particle of a statisticallyisotropic shape is defined as the mass of the particle divided by theminimum sphere envelope volume within which it can be enclosed. Featureswhich can contribute to low tap density include irregular surfacetexture and porous structure. Tap density can be measured by usinginstruments known to those skilled in the art such as the Dual PlatformMicroprocessor Controlled Tap Density Tester (Vankel, N.C.), a GeoPyc™instrument (Micrometries Instrument Corp., Norcross, Ga.), or SOTAX TapDensity Tester model TD2 (SOTAX Corp., Horsham, Pa.). Tap density can bedetermined using the method of USP Bulk Density and Tapped Density,United States Pharmacopia convention, Rockville, Md., 10^(th)Supplement, 4950-4951, 1999.

Fine particle fraction (FPF) can be used as one way to characterize theaerosol performance of a dispersed powder. Fine particle fractiondescribes the size distribution of airborne respirable dry particles.Gravimetric analysis, using a Cascade impactor, is one method ofmeasuring the size distribution, or fine particle fraction, of airbornerespirable dry particles. The Andersen Cascade Impactor (ACI) is aneight-stage impactor that can separate aerosols into nine distinctfractions based on aerodynamic size. The size cutoffs of each stage aredependent upon the flow rate at which the ACI is operated. The ACI ismade up of multiple stages consisting of a series of nozzles (i.e., ajet plate) and an impaction surface (i.e., an impaction disc). At eachstage an aerosol stream passes through the nozzles and impinges upon thesurface. Respirable dry particles in the aerosol stream with a largeenough inertia will impact upon the plate. Smaller respirable dryparticles that do not have enough inertia to impact on the plate willremain in the aerosol stream and be carried to the next stage. Eachsuccessive stage of the ACI has a higher aerosol velocity in the nozzlesso that smaller respirable dry particles can be collected at eachsuccessive stage.

If desired, a two-stage collapsed ACI can also be used to measure fineparticle fraction. The two-stage collapsed ACI consists of only the toptwo stages of the eight-stage ACI and allows for the collection of twoseparate powder fractions. Specifically, a two-stage collapsed ACI iscalibrated so that the fraction of powder that is collected on stage oneis composed of respirable dry particles that have an aerodynamicdiameter of less than 5.6 μm and greater than 3.4 μm. The fraction ofpowder passing stage one and depositing on a collection filter is thuscomposed of respirable dry particles having an aerodynamic diameter ofless than 3.4 μm. The airflow at such a calibration is approximately 60L/min. Formulation produced by the methods described herein can beeffectively delivered at airflow rates ranging from about 20 L/min toabout 60 L/min.

An ACI can be used to approximate the emitted dose, which herein iscalled gravimetric recovered dose and analytical recovered dose.“Gravimetric recovered dose” is defined as the ratio of the powderweighed on all stage filters of the ACI to the nominal dose. “Analyticalrecovered dose” is defined as the ratio of the powder recovered fromrinsing all stages, all stage filters, and the induction port of the ACIto the nominal dose. The FPF TD (<5.0) is the ratio of the interpolatedamount of powder depositing below 5.0 μm on the ACI to the nominal dose.The FPF RD (<5.0) is the ratio of the interpolated amount of powderdepositing below 5.0 μm on the ACI to either the gravimetric recovereddose or the analytical recovered dose.

Another way to approximate emitted dose is to determine how much powderleaves its container, e.g. capsule or blister, upon actuation of a drypowder inhaler (DPI). This takes into account the percentage leaving thecapsule, but does not take into account any powder depositing on theDPI. The emitted dose is the ratio of the weight of the capsule with thedose before inhaler actuation to the weight of the capsule after inhaleractuation. This measurement can also be called the capsule emittedpowder mass (CEPM).

A Multi-Stage Liquid Impinger (MSLI) is another device that can be usedto measure particle size distribution or fine particle fraction. TheMulti-stage liquid Impinger operates on the same principles as the ACI,although instead of eight stages, MSLI has five. Additionally, each MSLIstage consists of an ethanol-wetted glass frit instead of a solid plate.The wetted stage is used to prevent particle bounce and re-entrainment,which can occur when using the ACI. U.S. Pat. No. 8,614,255.

The subject technology also relates to a respirable dry powder orrespirable dry particles produced using any of the methods describedherein.

The dry particles of the subject technology can also be characterized bythe chemical stability of the salts or the excipients that the dryparticles comprise. The chemical stability of the constituent salts canaffect important characteristics of the particles including shelf-life,proper storage conditions, acceptable environments for administration,biological compatibility, and effectiveness of the salts. Chemicalstability can be assessed using techniques well known in the art. Oneexample of a technique that can be used to assess chemical stability isreverse phase high performance liquid chromatography (RP-HPLC).

If desired, the dry particles and dry powders described herein can befurther processed to increase stability. An important characteristic ofpharmaceutical dry powders is whether they are stable at differenttemperature and humidity conditions. Unstable powders will absorbmoisture from the environment and agglomerate, thus altering particlesize distribution of the powder.

Excipients, such as maltodextrin, may be used to create more stableparticles and powders. The maltodextrin may act as an amorphous phasestabilizer and inhibit the components from converting from an amorphousto crystalline state. Alternatively, a post-processing step to help theparticles through the crystallization process in a controlled way (e.g.,on the baghouse at elevated humidity) can be employed with the resultantpowder potentially being further processed to restore theirdispersibility if agglomerates formed during the crystallizationprocess, such as by passing the particles through a cyclone to breakapart the agglomerates. Another possible approach is to optimize aroundprocess conditions that lead to manufacturing particles that are morecrystalline and therefore more stable. Another approach is to usedifferent excipients, or different levels of current excipients toattempt to manufacture more stable forms of the salts.

The respirable dry particles and dry powders described herein aresuitable for inhalation therapies. The respirable dry particles may befabricated with the appropriate material, surface roughness, diameter,and tap density for localized delivery to selected regions of therespiratory system such as the deep lung or upper or central airways.For example, higher density or larger respirable dry particles may beused for upper airway delivery, or a mixture of varying size respirabledry particles in a sample, provided with the same or a differentformulation, may be administered to target different regions of the lungin one administration.

In order to relate the dispersion of powder at different inhalation flowrates, volumes, and from inhalers of different resistances, the energyrequired to perform the inhalation maneuver can be calculated.Inhalation energy can be calculated from the equation E=R²Q²V where E isthe inhalation energy in Joules, R is the inhaler resistance inkPa^(1/2)/LPM, Q is the steady flow rate in L/min and V is the inhaledair volume in L.

Healthy adult populations are predicted to be able to achieve inhalationenergies ranging from 2.9 to 22 Joules by using values of peakinspiratory flow rate (PIFR) measured by Clarke et al. (Journal ofAerosol Med, 6(2), p. 99-110, 1993) for the flow rate Q from two inhalerresistances of 0.02 and 0.055 kPa½/LPM, with a inhalation volume of 2 Lbased on both FDA guidance documents for dry powder inhalers and on thework of Tiddens et al. (Journal of Aerosol Med, 19, (4), p. 456-465,2006) who found adults averaging 2.2 L inhaled volume through a varietyof DPIs.

Dry powder particles can also be prepared using cone-jet mode ofelectrohydrodynamic atomization, as described by Li et al., ChemicalEngineering Science 61 (2006) 3091-3097. For example, an acetylsalicylicacid solution flowing through a needle can be subjected to an electricfield to generate droplets. The method is said to generate anear-monodispersed distribution of droplet relics, leading to formacetylsalicylic acid particulate crystals.

7. Methods of Treatment

In other aspects, the subject technology is a method for treating(including prophylactic treatment or reducing the risk) of acardiovascular disease (such as thrombosis), comprising administering tothe respiratory tract of a subject in need thereof an effective amountof respirable dry particles or dry powder, as described herein.

Cardiovascular diseases include, for example, atherosclerosis, coronaryartery disease (CAD), angina pectoris (commonly known as “angina”),thrombosis, ischemic heart disease, coronary insufficiency, peripheralvascular disease, myocardial infarction, cerebrovascular disease (suchas stroke), transient ischemic attack, arteriolosclerosis, small vesseldisease, elevated cholesterol, intermittent claudication orhypertension.

The respirable dry particles and dry powders can be administered to therespiratory tract of a subject in need thereof using any suitablemethod, such as instillation techniques, and/or an inhalation device,such as a dry powder inhaler (DPI) or metered dose inhaler (MDI). Anumber of DPIs are available, such as, the inhalers disclosed is U.S.Pat. Nos. 4,995,385 and 4,069,819, Spinhaler® (Fisons, Loughborough,U.K.), Rotahalers®, Diskhaler® and Diskus® (GlaxoSmithKline, ResearchTriangle Technology Park, N.C.), FlowCapss®, XCaps (Hovione, Loures,Portugal), Inhalators® (BoehringerIngelheim, Germany), Aerolizer®(Novartis, Switzerland), and others known to those skilled in the art.

Generally, inhalation devices (e.g., DPIs) are able to deliver a maximumamount of dry powder or dry particles in a single inhalation, which isrelated to the capacity of the blisters, capsules (e.g. size 000, 00,OE, 0, 1, 2, 3, and 4, with respective volumetric capacities of 1.37 ml,950 μl, 770 μl, 680 μl, 480 μl, 360 μl, 270 μl, and 200 μl) or othermeans that contain the dry particles or dry powders within the inhaler.Accordingly, delivery of a desired dose or effective amount may requiretwo or more inhalations. Preferably, each dose that is administered to asubject in need thereof contains an effective amount of respirable dryparticles or dry powder and is administered using no more than about 4inhalations. For example, each dose of respirable dry particles or drypowder can be administered in a single inhalation or 2, 3, or 4inhalations. The respirable dry particles and dry powders are preferablyadministered in a single, breath-activated step using a breath-activatedDPI. When this type of device is used, the energy of the subject'sinhalation both disperses the respirable dry particles and draws theminto the respiratory tract.

The respirable dry particles or dry powders can be delivered byinhalation to a desired area within the respiratory tract, as desired.It is well known that particles with an MMAD of about 1 μm to about 3μm, can be effectively delivered to the deep lung regions such as thealveolar spaces. Larger aerodynamic diameters, for example, from about 3μm to about 5 μm can be delivered to the central and upper airways.

For dry powder inhalers, oral cavity deposition is dominated by inertialimpaction and so characterized by the aerosol's Stokes number (DeHaan etal. Journal of Aerosol Science, 35 (3), 309-331, 2003). For equivalentinhaler geometry, breathing pattern and oral cavity geometry, the Stokesnumber, and so the oral cavity deposition, is primarily affected by theaerodynamic size of the inhaled powder. Hence, factors that contributeto oral deposition of a powder include the size distribution of theindividual particles and the dispersibility of the powder. If the MMADof the individual particles is too large, e.g. above 5 μm, then anincreasing percentage of powder will deposit in the oral cavity.Likewise, if a powder has poor dispersibility, it is an indication thatthe particles will leave the dry powder inhaler and enter the oralcavity as agglomerates. Agglomerated powder will perform aerodynamicallylike an individual particle as large as the agglomerate, therefore evenif the individual particles are small (e.g., MMAD of about 5 μm orless), the size distribution of the inhaled powder may have an MMAD ofgreater than about 5 μm, leading to enhanced oral cavity deposition.

Certain embodiments provide a powder in which the particles are small(e.g., MMAD of m or less, e.g. between about 1 μm to 5 μm), and arehighly dispersible (e.g. ¼ bar or alternatively, 0.5/4 bar of 2.0, andpreferably less than 1.5). The respirable dry powder may be comprised ofrespirable dry particles with an MMAD between 1 to 4 μm or 1 to 3 μm,and have a ¼ bar less than 1.4, or less than 1.3, and more preferablyless than 1.2.

The absolute geometric diameter of the particles measured at 1 bar usingthe HELOS system is not critical provided that the particle's envelopedensity is sufficient such that the MMAD is in one of the ranges listedabove, wherein MMAD is VMGD times the square root of the envelopedensity (MMAD=VMGD*sqrt (envelope density)). If it is desired to delivera high unit dose of salt using a fixed volume-dosing container, then,particles of higher envelop density are desired. High envelope densityallows for more mass of powder to be contained within the fixedvolume-dosing container. Preferable envelope densities are greater than0.1 g/cm³, greater than 0.25 g/cm³, greater than 0.4 g/cm³, greater than0.5 g/cm³, and greater than 0.6 g/cm³.

The respirable dry powders and particles of the subject technology canbe employed in compositions suitable for drug delivery via therespiratory system. For example, such compositions can include blends ofthe respirable dry particles of the subject technology and one or moreother dry particles or powders, such as dry particles or powders thatcontain another active agent, or that consist of or consist essentiallyof one or more pharmaceutically acceptable excipients.

Respirable dry powders and dry particles suitable for use in the methodsof the subject technology can travel through the upper airways (i.e.,the oropharynx and larynx), the lower airways, which include the tracheafollowed by bifurcations into the bronchi and bronchioli, and throughthe terminal bronchioli which in turn divide into respiratory bronchioleleading then to the ultimate respiratory zone, the alveoli or the deeplung. In one embodiment of the subject technology, most of the mass ofrespirable dry powders or particles deposit in the deep lung. In anotherembodiment of the subject technology, delivery is primarily to thecentral airways. In another embodiment, delivery is to the upperairways.

The respirable dry particles or dry powders of the subject technologycan be delivered by inhalation at various parts of the breathing cycle(e.g., laminar flow at mid-breath). An advantage of the highdispersibility of the dry powders and dry particles of the subjecttechnology is the ability to target deposition in the respiratory tract.For example, breath controlled delivery of nebulized solutions is arecent development in liquid aerosol delivery (Dalby et al. inInhalation Aerosols, edited by Hickey 2007, p. 437). In this case,nebulized droplets are released only during certain portions of thebreathing cycle. For deep lung delivery, droplets are released in thebeginning of the inhalation cycle, while for central airway deposition,they are released later in the inhalation.

The dry powders of this subject technology provide advantages fortargeting the timing of drug delivery in the breathing cycle and alsolocation in the human lung. Because the respirable dry powders of thesubject technology can be dispersed rapidly, such as within a fractionof a typical inhalation maneuver, the timing of the powder dispersal canbe controlled to deliver an aerosol at specific times within theinhalation.

With a highly dispersible powder, the complete dose of aerosol can bedispersed at the beginning portion of the inhalation. While thepatient's inhalation flow rate ramps up to the peak inspiratory flowrate, a highly dispersible powder will begin to disperse already at thebeginning of the ramp up and could completely disperse a dose in thefirst portion of the inhalation. Since the air that is inhaled at thebeginning of the inhalation will ventilate deepest into the lungs,dispersing the most aerosol into the first part of the inhalation ispreferable for deep lung deposition. Similarly, for central deposition,dispersing the aerosol at a high concentration into the air which willventilate the central airways can be achieved by rapid dispersion of thedose near the mid to end of the inhalation. This can be accomplished bya number of mechanical and other means such as a switch operated bytime, pressure or flow rate that diverts the patient's inhaled air tothe powder to be dispersed only after the switch conditions are met.

Aerosol dosage, formulations and delivery systems may be selected for aparticular therapeutic application, as described, for example, in Gonda,I. “Aerosols for delivery of therapeutic and diagnostic agents to therespiratory tract,” in Critical Reviews in Therapeutic Drug CarrierSystems, 6: 273-313 (1990); and in Moren, “Aerosol Dosage Forms andFormulations,” in Aerosols in Medicine, Principles, Diagnosis andTherapy, Moren, et al., Eds., Esevier, Amsterdam (1985).

Suitable intervals between doses that provide the desired therapeuticeffect can be determined based on the severity of the condition, overallwell-being of the subject and the subject's tolerance to respirable dryparticles and dry powders and other considerations. Based on these andother considerations, a clinician can determine appropriate intervalsbetween doses. Respirable dry particles and dry powders may beadministered once, twice or three times a day or on an as needed basis.

In various embodiments the amount of NSAID, such as acetylsalicyclicacid, delivered to the respiratory tract (e.g., lungs, respiratoryairway) is about 0.001 mg/kg body weight/dose to about 2 mg/kg bodyweight/dose, about 0.002 mg/kg body weight/dose to about 2 mg/kg bodyweight/dose, about 0.005 mg/kg body weight/dose to about 2 mg/kg bodyweight/dose, about 0.01 mg/kg body weight/dose to about 2 mg/kg bodyweight/dose, about 0.02 mg/kg body weight/dose to about 2 mg/kg bodyweight/dose, about 0.05 mg/kg body weight/dose to about 2 mg/kg bodyweight/dose, about 0.075 mg/kg body weight/dose to about 2 mg/kg bodyweight/dose, about 0.1 mg/kg body weight/dose to about 2 mg/kg bodyweight/dose, about 0.2 mg/kg body weight/dose to about 2 mg/kg bodyweight/dose, about 0.5 mg/kg body weight/dose to about 2 mg/kg bodyweight/dose, or about 0.75 mg/kg body weight/dose to about 2 mg/kg bodyweight/dose.

In certain embodiments, at least about 50%, at least about 60%, at leastabout 70%, at least about 75%, at least about 80%, at least about 85%,at least about 90%, at least about 95%, or at least about 99%, of theadministered acetylsalicylic acid reaches the systemic circulation of asubject within about 60 minutes upon administration, or within about 40minutes upon administration, or within about 30 minutes uponadministration, or within about 20 minutes upon administration, orwithin about 15 minutes upon administration, or within about 5 minutesupon administration.

The dosing of acetylsalicyclic acid may be adjusted so that PGI2synthesis capacity of the nasal, bronchial or pulmonary epithelial ofendothelial cells, including nasal mucosa cells, is not inhibited.

In certain embodiments, the method and delivery devices described hereincan deliver acetylsalicylic acid, and pharmacologically active metabolicbyproducts of acetylsalicylic acid thereof, to the systemic circulation,at levels that are substantially the same, or higher as compared tothose delivered by oral administration of about 30 mg of acetylsalicylicacid, about 40 mg of acetylsalicylic acid, about 50 mg ofacetylsalicylic acid, about 80 mg of acetylsalicylic acid or about 160mg of acetylsalicylic acid.

The doses of acetylsalicylic acid administered in order to achieve alevel (or an average level among a population of patients) that issubstantially the same, or higher as compared to those delivered by oraladministration of about 30 mg, about 40 mg, about 50 mg, about 80 mg, orabout 160 mg of acetylsalicylic acid can be determined by conventionalmethods. The dosing, administration techniques and schedules are knownin the art and are within the ability of the skilled clinician. Forexample, the serum level of acetylsalicylic acid, or a metabolitethereof, in a subject (or average serum level among a population ofsubjects) can be determined by conventional pharmacokinetic orpharmacodynamics studies.

In certain embodiments, the method and delivery devices described hereincan deliver acetylsalicylic acid to the systemic circulation such thatthe circulating plasma level of acetylsalicylic acid is at least about 1μg/mL, at least about 2 μg/mL, at least about 3 μg/mL, at least about 4μg/mL, at least about 5 μg/mL, oat least about 6 μg/mL, within about 60minutes upon administration, or within about 40 minutes uponadministration, or within about 30 minutes upon administration, orwithin about 20 minutes upon administration, or within about 15 minutesupon administration, or within about 5 minutes upon administration.

In other embodiments, the method and delivery devices described hereincan deliver acetylsalicylic acid to the systemic circulation such thatcirculating plasma level of salicylate is about 8 μg/mL, about 9 μg/mL,about 10 μg/mL, about 11 μg/mL, about 12 μg/mL, or about 15 g/mL, withinabout 60 minutes upon administration, or within about 40 minutes uponadministration, or within about 30 minutes upon administration, orwithin about 20 minutes upon administration, or within about 15 minutesupon administration, or within about 5 minutes upon administration.

If desired or indicated, the respirable dry particles and dry powdersdescribed herein can be administered with one or more other therapeuticagents. The other therapeutic agents can be administered by any suitableroute, such as orally, parenterally (e.g., intravenous, intraarterial,intramuscular, or subcutaneous injection), topically, by inhalation(e.g., intrabronchial, intranasal or oral inhalation, intranasal drops),rectally, vaginally, and the like. The respirable dry particles and drypowders can be administered before, substantially concurrently with, orsubsequent to administration of the other therapeutic agent. Preferably,the respirable dry particles and dry powders and the other therapeuticagent are administered so as to provide substantial overlap of theirpharmacologic activities.

The following examples of specific aspects for carrying out theembodiments of present invention are offered for illustrative purposesonly, and are not intended to limit the scope of the present inventionin any way.

Example 1 Development of Acetylsalicylic Acid Particles for Inhalation

This study was designed to develop phospholipid coated acetylsalicylicacid particles with particle size less than 2.0 μm for deep lung tissuedelivery. This development work was carried out to achieve followingtarget particle size: Dv50 of 0.5 nm to 2.0 μm; Dv90 of 1.5 to 2.0 μm.

Jet milling was selected as a method for micronization ofacetylsalicylic acid particles to achieve target particle size. Jetmilling operation was successfully reproduced with Dv50s of 0.4 m andDv90s of 1.3 μm and 1.6 μm for the two batches manufactured. Micronizedparticles were then spray-dried using DSPC(1,2-distearoyl-(sn)-glycero-3-phosphocholine) or soy lecithin to reduceparticle agglomeration and irritation on inhalation. 79% yield forDSPC/acetylsalicylic acid and 54% yield for soy lecithin/acetylsalicylicacid were obtained.

Particle size analysis at each step was carried out. Spray-driedDSPC/acetylsalicylic acid particles ranged from 1.8 to 3.6 μm andlecithin/acetylsalicylic acid particles ranged from 1.7 to 3.3 μm.

DSC studies showed no change in the crystalline structure ofacetylsalicylic acid before and after spray-drying with DSPC. TGA showed0.6% moisture content for both pre- and post-spray-dried particlesindicating absence of any residual solvent after spray-drying.

Formulation Development

1. Acetylsalicylic Acid

Rhodine 3040 US obtained from Rhodia Inc. was used for all experiments.Particles were dispersed in 0.1% w/w docusate sodium in water andobserved under a light microscope to confirm particle size. Particlesranging from 66 to 280 μm were observed confirming data from certificateof analysis, though observed “rounding” of the particulates isindicative of partial dissolution in the aqueous dispersant.

2. Particle Size Analysis

Particle size analysis was carried out using laser diffraction and lightmicroscopy.

2.1. Laser Diffraction

Horiba LA-950 V2 with fraction cell was used for laser diffractionstudies using the following parameters: dispersion media: 0.05% w/w soylecithin dissolved in n-hexane; refractive index of media: 1.334;refractive index of acetylsalicylic acid particles: 1.5623; i-value:0.01. The i-value is an imaginary component that is used by the laserdiffraction algorithm to account for the absorption of light by theparticles. A stock dispersion of particles in the same media wasprepared and added drop-wise to the fraction cell containing a magneticstirrer bar until the intensity meter showed red laser between 80% and90%, while blue laser was between 70% and 90%. Once stabilized, thevolumetric mean diameter Dv10, Dv50 and Dv90 were measured. This laserdiffraction method developed for uncoated particles was not used forspray-dried phospholipid/acetylsalicylic acid particles as they did notdisperse well in the selected media.

2.2 Light Microscopy

Photomicrographs of the pre- and post-micronized uncoated particles weretaken by dispersing them in a solution of 0.1% w/w docusate sodium inpurified water USP, and using a digital imaging light microscope(Olympus BX51 with Clemex ST-2000 controller) at 400-times or 1000-timesmagnification. As spray-dried phospholipid/acetylsalicylic acidparticles were found to not disperse well in the selected media,photomicrographs were taken after spreading them in the dry state overglass slides.

3. Jet Milling Trials using Sturtevant Qualification Mill

Initial work was carried out using a Sturtevant Qualification mill withventuri #1 using nitrogen as the carrier gas. Material was fed throughvibratory feeder at controlled rate and at predetermined feed and grindpressure. The effect of grinding pressure, feed rate, and second passwere studied on particle size reduction and the conditions are reportedin Table 4.

TABLE 4 Jet Milling Trials using Sturtevant Qualification MillFormulation 3694 3695 3701 3702 Milling run Pass#1 Pass#1 Pass#1 Pass#2of Formulation 3695 P_Feed 7 7 7 7 (bar) P_Grind 3.5 5 3.5 5 (bar)F_Flow 17 17 54 7 (g/hr)3.1 Effect of Grind Pressure

Formulations 3694 and 3695 were compared to study effect of grindpressure on PSD (particle size distribution). Laser diffraction andmicroscopy results were obtained are presented in Table 5. Microscopyand laser diffraction data were found to correlate very well. Whengrinding pressure was increased from 3.5 to 5 bar, a measurable decreasein particle size was observed as would be expected.

TABLE 5 Effect of Grind Pressure on Acetylsalicylic acid Particle SizeAverage (% RSD)/n = 3 Fomnflation 3694 3695 Dv10 (μm) 1.9 (1.0) 1.2(2.2) Dv50 (μm) 3.3 (1.8) 2.4 (1.9) Dv90 (μm) 5.9 (2.8) 4.9 (2.6)Microscopy 1.7-5.1 2.0-3.5 (μm)3.2 Effect of Feed Rate

Formulations 3694 and 3701 were compared to study the effect of materialfeed rate on particle size using microscopy (Table 6). Clearly, as theflow rate was increased from 17 g/hr to 54 g/hr, significantly largerparticles were obtained. This is likely the result of new materialentering the milling chamber and pushing out particles to the collectionbag before they undergo sufficient attrition.

TABLE 6 Effect of Feed Rate on Particle Size Formulation 3694 3701Microscopy 1.7-5.1 5.2-42.1 (μm)3.3 Effect of Second Milling Pass

In order to achieve the target particle size of Dv50 of 1.5 μm and Dv90of 2 μm, formulation 3695 was passed the through mill for a second pass.Particle size analysis was carried out using laser diffraction andmicroscopy (Table 7). Significant particle size reduction was achievedon the second pass through the jet mill which implies thatacetylsalicylic acid particles undergo first order size reduction, andthat final particle size obtained depends upon initial particle sizeused.

TABLE 7 Effect of Second Milling Pass on Particle Size Formuiation 36953702 Microscopy 2.0-3.5 0.8-2.4 (μM)4. Jet Milling Using Sturtevant Sanitary Design Mill

In order to achieve higher feed rate with better control as well as toincrease batch size, the larger 2″ sanitary design mill was usedaccording to parameters listed in Table 8. Material was processed on asecond pass as well to reduce particle size to target. Formulations 3727and 3734 were compared with 3705 and 3725 processed using theQualification mill respectively to study reproducibility in PSD. Anantistatic device was necessary to feed the powder for the second passto minimize the effects of the static electricity imparted during thefirst pass.

TABLE 8 Jet Milling using Sturtevant Sanitary Design Mill Mill UsedQ-Mill 2° Mill Formulation 3705 3725 3727 3734 Milling run Pass#1 Pass#2Pass#1 Pass#2 (Formulation (Formulation 3705) 3727) P_Venturi (bar) 4.12.8 4.1 2.9 P_Grind (bar) 2.8 2.1 2.8 2.1 F_Flow (g/hr) 132 78 142 59Batch size (g) 80 50 200 120

Aggregated particles with high static charge were obtained in all cases.

4.1 Particle Size Analysis

Particle size analysis of above formulations was carried out using laserdiffraction and microscopy (Table 9, FIG. 1 and FIG. 2). Reproducibleresults in particle size reduction were obtained with comparable Dv10,Dv50 and Dv90 values between the two mill sizes, even with batch sizeincreased from 80 g to 200 g for the first pass and from 50 g to 120 gfor the second pass. Monomodal PSD was obtained for the first pass whilea bimodal distribution was observed for the second pass.

TABLE 9 Particle Size Analysis of Jet Milled Acetylsalicylic AcidFormulations Prepared Using Sanitary Design Mill Average (% RSD)/n = 3Formulation 705 325 327 3734 Dv10 (μm) 0.9 (5.2) 0.1 (1.9)  0.8 (10.0)0.1 (1.6) Dv50 (μm) 1.5 (3.3) 0.4 (6.3) 1.3 (5.2)  0.4 (12.9) Dv90 (μm)2.6 (4.2) 1.8 (6.0) 2.2 (6.7) 1.5 (5.1) Microscopy 1.2-2.6 μm 0.9-1.8 μm1.1-3.2 μm 0.9-2.3 μm5. Coating

Spray-drying was used for coating. Jet milled formulation 3734,processed two passes on the 2″ sanitary mill, was used further to coatwith either DSPC or soy lecithin. Particles were dispersed in n-hexanecontaining lipid and spray-drying was selected as a method to removesolvent. In order to achieve coating around all individual particles, itwas required to disperse Jet milled particles completely withoutsettling, and therefore, continuous stirring was employed throughout thespray-drying operation.

5% w/w DSPC was used as it was found from previous work to mitigateirritation when inhaled. Additionally, soy lecithin was also used in theconcentration of 0.1% w/w. As acetylsalicylic acid is insoluble inn-hexane, it was selected as a dispersion media for the micronizedparticles. Also, it has boiling point of 70° C. which is much below themelting point of acetylsalicylic acid (˜135° C.) and therefore, an inlettemperature of 85° C. should remove solvent without affecting theacetylsalicylic acid particles.

A Buchi-290 spray dryer equipped with nozzle of 0.7 mm diameter was usedfor the study. Spray-drying was performed using nitrogen as the carriergas with the aspirator set at 100% capacity. The flow rate of nitrogenwas adjusted to 1052 L/hr (50 mm in rotameter). Before feeding the stockdispersion, feed rate was adjusted using dispersing media alone toachieve targeted outlet temperature and stabilization of the system.

5.1 Spray-Drying Using DSPC

DSPC (Lipoid PC 18:0/18:0) is an endogenous lung phospholipid with aphase transition temperature of 55° C. On heating at this temperature,DSPC transforms into a liquid crystalline phase from the gel phase, andthe phospholipid layer is dispersed in n-hexane as a monolayer with arandom and non-rigid structure. When jet milled acetylsalicylic acidparticles are dispersed in the DSPC/Hexane solution, a well dispersedcolloidal suspension was formed without noticeable settling. From this,it was hypothesized that spray-drying should be able to coat individualacetylsalicylic acid particles on solvent removal. Details of theprocessing are reported in Table 10.

TABLE 10 Spray-Drying Parameters for DSPC/Acetylsalicylic acidFormulation Formulation 3739 Suspension preparation DSPC (g) 050n-hexane (g) 490 Jet milled ASA (g) 9.50 % Soild in feed 2 Suspension 55temperature (°C.) Spray-drying parameters Inlet temperature 85 (°C.)Outlet temperature 56 (°C.) Flow rate (g/min) 3.9 Flow meter (mm) 50Suspension 55 temperature (°C.) % Yield 79

No excessive sticking to the spray-drying chamber was observed duringprocessing and a yield of 79% was obtained. Also, the coated particlesobtained were observed to be denser and less static than uncoatedparticles.

5.1.1 Particle Size Analysis

The spray-dried DSPC coated particles were found to not disperse as wellin the 0.05% w/w soy lecithin/n-hexane solution used for particle sizeanalysis of the uncoated acetylsalicylic acid particles. Someagglomeration was observed by microscopy compared to uncoated, thoughPSD ranges of the primary particles was collated from the microscopicimages (Table 11).

TABLE 11 Particle Size of Micronized Uncoated and Spray-Dried DSPC/Acetylsalicylic acid Particles Formulation 3734 3739 MicronizedSpray-dried Description uncoated DSPC/aspirin Microscopy 0.9-2.3 1.8-3.6(μm)5.1.2 Differential Scanning Calorimetry (DSC)

A DSC study was carried out on raw acetylsalicylic acid, uncoated milledparticles of formulation 3734 and spray-dried DSPC/acetylsalicylic acidparticles of formulation 3739 to study any change in the crystallinityof the acetylsalicylic acid induced from processing.

Samples were sealed in 40 μL aluminum pans with pierced lids andanalyzed using a differential scanning calorimeter (Mettler-Toledo DSCequipped with STAR® software V10.00). The samples were heated from 25°C. to 160° C. at a rate of 10° C. per minute. An empty pan served as thereference.

In all samples, a sharp endothermic peak corresponding toacetylsalicylic acid melting was observed. No other polymorphicconversion was observed. Also, no significant shift in peak was observedconfirming no change in crystallinity of the acetylsalicylic acid onprocessing (FIG. 3, Table 12).

TABLE 12 DSC Analysis of Raw, Micronized Uncoated and Spray-DriedDSPC/Acetylsalicylic acid Particles Onset Peak temperature temperatureSample (°C.) (°C.) Rhodine 3040 142.5 144.7 US raw Micronized 141.6142.8 uncoated aspirin Spray-dried 139.7 141.3 DSPC/aspirin5.1.3 Thermogravimetric Analysis (TGA)

TGA was carried out for the micronized uncoated acetylsalicylic acidparticles of formulation 3734 and spray-dried DSPC/acetylsalicylic acidparticles of formulation 3739 to evaluate those for residual solvent andchange in moisture content of the particles on spray-drying.

TGA of spray-dried powder carried out in 40 μL aluminum open pans byheating them from 25° C. to 160° C. at a rate of 10° C. per minute usingMettler-Toledo TGA/DSC1 equipped with STAR® software V10.00. The %weight loss was measured from 25° C. to 120° C. and compared betweenpre- and post-spray-drying.

TGA data suggests that there is no residual hexane in spray-driedparticles, as the % weight loss before and after spray drying showsimilar values. The 0.57% weight loss is likely indicative of themoisture content of pre- and post-spray-dried acetylsalicylic acidparticles (FIG. 4, Table 13).

TABLE 13 % Weight Loss for Micronized Uncoated and Spray-Dried DSPC/Acetylsalicylic acid Particles Formulation % weight toss (g) Micronizeduncoated 0.58 aspirin Spray-dried 0.57 DSPC/aspirin5.2 Spray-Drying Acetylsalicylic Acid Particles Using Soy Lecithin

Soy lecithin was selected as an excipient as it is also approved forinhalation drug delivery and it was able to disperse jet milledacetylsalicylic acid particles well. Therefore, it was expected to beable to coat individual acetylsalicylic acid particle on solventremoval. Soy lecithin was dissolved in n-hexane and jet milledacetylsalicylic acid particles dispersed in it with stirring. However,unlike the dispersion of acetylsalicylic acid in DSPC, the soy lecithinin 0.1% w/w concentration was not able to form colloidal dispersion, andsome settling was observed. Therefore, continuous stirring of the feedsuspension during spray-drying was used to maintain the dispersion ofthe acetylsalicylic acid particles. Spray-drying was carried out toremove n-hexane and coat acetylsalicylic acid particles using theparameters in Table 14. A 54% yield was obtained.

TABLE 14 Spray-Drying Parameters for Soy Lecithin/Acetylsalicylic acidFormulation Formulation 3740 Suspension preparation Soy lecithin (g)0.01 n-hexane (g) 490 Jet milled ASA (g) 9.99 % Solid in feed 2Suspension RT temperature (°C.) Spray-drying parameters Inlettemperature 85 (°C.) Outlet temperature 59 (°C.) Flow rate (g/min) 3.9Flow meter (mm) 50 Suspension RT temperature Yield 54%5.2.1 Particle Size Analysis

Particle size analysis was carried out using powder microscopy andcompared with micronized uncoated and spray-dried DSPC/acetylsalicylicacid. Particle size of both spray-dried formulations suggestsatisfactory results (Table 15).

TABLE 15 Particle Size Analysis of Spray-dried SoyLecithin/Acetylsalicylic acid Particles Formulation 3734 3739 3740Micronized Spray-dried Spray-dried uncoated DSPC/ soy Descriptionaspirin aspirin lecithin/Aspirin Microscopy 0.9-2.3 1.8-3.6 1.7-3.3 (μm)Conclusions

The micronization of acetylsalicylic acid yielded an approximately 70fold reduction in the starting particle size. Spray-drying with DSPC orsoy lecithin resulted in satisfactory particle size for deep lung tissuedrug delivery with maximum size of 3.6 μm. Spray-driedDSPC/acetylsalicylic acid particles were found to be less static thansoy lecithin/acetylsalicylic acid particles, and even less static thanmicronized uncoated acetylsalicylic acid particles. The crystallinestructure of acetylsalicylic acid did not change during milling or spraydrying as observed by DSC study. DSC studies also suggested absence ofany other event such as polymorph conversion during processing. Notraces of residual solvent found in spray dried DSPC/acetylsalicylicacid during TGA analysis.

Example 2 Emitted Dose Analysis of DSPC/Acetylsalicylic Acid Particlesand Soy Lecithin/Acetylsalicylic Acid Particles

DPI devices, e.g. Plastiape, were used to evaluate emitted doses of theDSPC/acetylsalicylic acid particles and soy lecithin/acetylsalicylicacid particles.

TABLE 16 % Initial Final Total Emitted Flow Fill Device Device Emitted(based DPI Rate Weight Weight Weight Dose on fill Powder Type Device(slpm) (mg) (g) (g) (mg) weight) Acetylsalicylic Plastiape 100.0 20.1110.49661 10.48124 15.37 76.4 acid (Soy Lecithin) AcetylsalicylicPlastiape 100.0 32.81 10.55421 10.53301 21.20 64.6 acid (Soy Lecithin)

Lactose was used to test proper device setup.

Example 3 Particle Size Distribution (PSD) Analysis of InhaledAcetylsalicylic Acid by Dry Dispersion and Laser Diffraction

Particle size analysis was carried out using laser diffraction analysisof dry dispersed spray-dried DSPC/acetylsalicylic acid particles offormulation 3739 (Table 17), and spray-dried soylecithin/acetylsalicylic acid particles formulation 3740 (Table 18) (seeExample 1 for the preparation of DSPC/acetylsalicylic acid particles andsoy lecithin/acetylsalicylic acid particles).

TABLE 17 Primary Pressure Particle Size (μm) Optical Lens (bar)Replicate X10 X50 X90 VMD GSD Conc. R3 1.0 1 0.96 2.29 4.47 2.56 1.828.71 R2 1.0 1 0.83 2.31 4.49 2.56 1.90 7.56 2 0.79 2.24 4.44 2.49 1.937.10 R1 0.7 1 0.66 2.63 5.27 2.88 2.07 9.21 2 0.64 2.59 5.18 2.84 2.075.72 0.9 1 0.62 2.34 4.63 2.57 2.01 4.78 2 0.57 2.31 4.68 2.55 2.09 5.981.0 1 0.58 2.31 4.69 2.54 2.11 10.98 2 0.60 2.34 4.69 2.56 2.08 6.53 30.56 2.31 4.76 2.57 2.14 6.74 4 0.57 2.26 4.51 2.48 2.07 7.97 5 0.582.28 4.53 2.49 2.06 8.20 1.2 1 0.56 2.13 4.17 2.32 2.03 4.38 2 0.55 2.124.17 2.32 2.05 12.60 2.0 1 0.60 2.03 4.03 2.54 1.97 6.03 2 0.54 2.034.25 2.57 2.12 7.63 3.0 1 0.55 1.84 3.68 2.13 2.03 8.88 2 0.52 1.81 3.632.01 2.07 8.49 4.0 1 0.47 1.79 3.64 2.00 2.12 6.41 2 0.52 1.80 3.58 2.002.03 7.56

TABLE 18 Primary Pressure Particle Size (μm) Optical Lens (bar)Replicate X10 X50 X90 VMD GSD Conc. R1 1.0 1 0.50 1.91 3.90 2.12 2.1111.28 2 0.50 1.89 3.73 2.07 2.03 3.89 3 0.49 1.90 3.83 2.11 2.09 9.58 40.52 1.90 3.66 2.06 2.00 4.70 5 0.49 1.90 3.83 2.10 2.08 6.84 Average0.502 1.900 3.792 2.091 2.062 % RSD RSD: relative standard deviation.

Example 4 NGI (Next-Generation Impactor) Analysis of Spray DriedAcetylsalicylic Acid/DPSC Particles

The dry powders of Example 1 were evaluated for aerodynamic performance.The DPI device used was a monodose inhaler. The NGI test conditionsranged between 20° C. and 25° C., and between 40% and 50% RH (relativehumidity) (Table 19).

TABLE 19 NGI 1 NGI 2 NGI 3 NGI 4 NGI 5 Controlled 21.83 C./46.7% RH22.66 C./47.3% RH 21.93 C./46.9% RH 21.93 C./46.9% RH 21.99 C./43.1% RHcondition Measured 99.1 SLPM 98.4 SLPM 97.6 SLPM 100.0 SLPM 100.5 SLPMFlow

Table 20 shows the aerodynamic properties of DSPC/acetylsalicylic acidparticles.

TABLE 20 2 capsules 1 capsule NGI 1 NGI 2 NGI 3 NGI 4 NGI 5 Device, l μg7876.4 9010.6 4267.0 4118.0 5115.8 Capsule 1, μg 653.6 717.9 484.6 464.5670.5 Capsule 2, μg 616.2 560.3 NA NA NA Induction Port, 11611.6 14550.87253.0 7454.4 6792.6 μg Stage 1, μg 10232.0 9393.6 3704.8 4257.6 5481.2Stage 2, μg 17402.0 16198.0 8284.4 8136.4 8758.4 Stage 3, μg 10882.49993.6 5600.8 4976.4 5087.6 Stage 4, μg 4884.0 4864.4 2791.2 2387.22273.6 Stage 5, μg 1670.0 1514.8 983.2 757.6 891.2 Stage 6, μg 983.81076.6 619.8 471.9 530.3 Stage 7, μg 575.6 498.2 318.9 262.9 284.1 MOC,μg 320.8 292.4 134.0 158.0 201.7 Nozzles, μg 5364.8 6363.2 2546.4 2833.63280.8 Nominal loaded 74 74 37 37 37 mass (mg) ED (mg) 63.93 64.75 32.2431.70 33.58 Nominal % ED 86% 88% 87% 86% 91% (mg) FPD (mg) 32.2 30.816.6 15.1 16.0 FPF (%) 50.4 47.5 51.5 47.7 47.5 MMAD (μm) 3.94 3.93 3.623.91 4.12 GSD 1.91 1.94 1.91 1.94 2.00 Recovery (%) 99.8 100.0 101.2100.1 103.7

Example 5 NGI Analysis of Spray Dried Acetylsalicylic Acid/Soy LecithinParticles

The dry powders of Example 1 were evaluated for aerodynamic performance.The DPI device used was a monodose inhaler. The NGI test conditionsranged between 20° C. and 25° C., and between 40% and 50% RH (relativehumidity) (Table 21).

TABLE 21 NGI 1 NGI 2 NGI 3 NGI 4 NGI 5 Controlled 22.57 C./49.6 22.16C./48.7% 22.14 C./47.9% 21.76 C./45.1% 21.66 C./45.1% condition % RH RHRH RH RH Measured 98.7 SLPM 97.6 SLPM 99.0 SLPM 100.0 SLPM 97.5 SLPMFlow

Table 22 shows the aerodynamic properties of soylecithin/acetylsalicylic acid particles.

TABLE 22 2 capsules 1 capsule NGI 1 NGI 2 NGI 3 NGI 4 NGI 5 Device, μg13139.2 15032.8 7664.0 6554.6 8382.0 Capsule 1, μg 1259.1 1607.1 1595.11078.2 916.0 Capsule 2, μg 2893.7 1050.2 NA NA NA Induction Port, μg5834.4 5586.6 3008.0 3604.4 3795.8 Stage 1, μg 4378.4 5104.0 1962.02274.8 2266.0 Stage 2, μg 12060.0 12890.8 5726.0 6028.0 6028.0 Stage 3,μg 15818.4 16041.6 7544.0 7687.2 7712.0 Stage 4, μg 11276.8 11301.65556.8 5345.6 5485.6 Stage 5, μg 3305.2 3182.0 1692.4 1622.4 1694.4Stage 6, μg 1272.6 1161.2 749.5 728.2 658.7 Stage 7, μg 708.4 605.2436.4 414.9 366.2 MOC, μg 340.8 375.8 231.8 228.6 236.9 Nozzles, μg4105.6 4928.0 1812.8 2306.4 2121.6 Nominal loaded 74 74 37 37 37 mass(mg) ED (mg) 59.10 61.18 28.72 30.24 30.37 Nominal % ED 80% 83% 78% 82%82% (mg) FPD (mg) 42.7 43.7 20.9 21.2 21.2 FPF (%) 72.3 71.5 72.7 70.069.7 MMAD (pm) 2.71 2.79 2.65 2.72 2.72 GSD 1.72 1.73 1.75 1.75 1.75Recovery (%) 104.6 104.0 104.4 103.1 104.5

HPLC analysis of acetylsalicylic acid in NGI and Delivered Dose sampleswas carried out as follows.

Equipment

The HPLC column was Phenomenex Luna C18(2) 5 μm, 4.6×100 mm. ShimadzuHPLC Equipment was used, including Shimadzu SIL-HTC Autosampler,Shimadzu CTO-10ASVP Column Oven, Shimadzu LC-10ADVP Binary HPLC Pump,Shimadzu DGU-14A Inline Degasser, Shimadzu UV Detector, and Computerwith Shimadzu Class VP software.

Materials

Mobile Phase A was 69:28:3 Water:Methanol:Glacial Acetic Acid. MobilePhase B was 97:3 Methanol:Glacial Acetic Acid. Diluent was 95:5Methanol:Glacial Acetic Acid. Needlewash was 50:50 Water:Methanol. Theworking standard was 750 μg/mL acetylsalicylic acid (working standard A“WSA” and working standard B “WSB”).

HPLC Conditions and Analysis

Flow rate was 2.0 mL/min. The sample injection volume was 10 μL. Thegradient was run according to the timing scheme in Table 23.

TABLE 23 HPLC Gradient Program Time (min) % B 0.00 0.0 3.80 0.0 3.81100.0 5.80 100.0 5.81 0.0 8.00 STOP

The analysis of the samples was in the following sequence:

A. Blank (2 injections) B. Working Standard A (6 injections) C. WorkingStandard B (2 injections) D. Blank (1 injection) E. Sample (1 injectioneach) F. WSB (QC Standard) (1 injection)

Repeat steps E-F as necessary ensuring that the last injection of asequence is a QC standard.

The standard agreement between WSA and WSB must be within 97.0-103.0%.The QC standard agreement between the ongoing standard analysis andinitial analysis (n=2) for WSB must between 97.0-103.0%.

The standard agreement between WSA and WSB was calculated according tothe equation below.

${SA} = {\frac{A_{WSA}}{A_{WSB}} \times \frac{C_{WSB}}{C_{WSA}} \times 100}$

-   -   Where:    -   SA=Standard Agreement (%)    -   A_(WSA)=WSA Average Area (n=6)    -   A_(WSB)=WSB Average Area (n=2)    -   C_(WSA)=WSA Theoretical Concentration (μg/mL)    -   C_(WSB)=WSB Theoretical Concentration (μg/mL)    -   100=Conversion to %

The % recovery of the QC standard(s) was calculated according to theequation below.

${QC} = {\frac{A_{QC}}{A_{WSB}} \times 100}$

-   -   Where:    -   QC=QC % Recovery    -   A_(QC)=QC Area    -   A_(WSB)=Initial WSB Average Area (n=2)    -   100=Conversion to %

The concentration of samples was calculated according to the equationbelow.

$C_{SX} = {\frac{A_{SX}}{A_{WSA}} \times C_{WSA}}$

-   -   Where:    -   C_(SX)=Sample Concentration (g/mL)    -   A_(SX)=Sample Area    -   A_(WSA)=WSA Average Area (n=6) Area    -   C_(WSA)=Theoretical WSA Concentration (g/mL)

Example 6 Spray Pattern and Plume Geometry

Spray pattern and plume geometry characterization of spray pattern andplume geometry of the formulation will be evaluated using standardmethodology (see, http://www.proveris.com/products/sprayview/; see also,http://www.oxfordlasers.com/imaging/spray-pattern-plume-geometry-measurement/).

Various factors can affect the spray pattern and plume geometry,including the size and shape of the actuator orifice, the design of theactuator, the size of the metering chamber, the size of the stem orificeof the valve, the vapor pressure in the container, and the nature of theformulation. Spray pattern testing will be tested on all formulationsunder a variety of different temperature and humidity conditions.

Example 7—Dry Powder Stability Testing

The following test parameters will be analyzed for the dry powderformulations.

i. Appearance and Color

The appearance of the content of the container and the appearance of thecontainer and closure system (i.e., the valve and its components and theinside of the container) will be analyzed. To determine whether there isany color is associated with the formulation (either present initiallyor from degradative processes occurring during shelf life.

ii. Microbial Limits

The microbial quality will be controlled for the total aerobic count,total yeast and mold count, and freedom from designated indicatorpathogens. Appropriate testing will be done to show that the drugproduct does not support the growth of microorganisms and that microbialquality is maintained throughout the expiration period.

iii. Water or Moisture Content

Water or moisture content will be analyzed. Changes in particle sizedistribution, morphic form, and other changes such as crystal growth oraggregation will be evaluated.

iv. Dose Content Uniformity

Dose content uniformity will be evaluated across multiple batches,formulations and under stability testing condition. The amount of activeingredient per determination will be evaluated to show that the emitteddose is 80-120 percent of label claim for more than one of tencontainers. The dose content uniformity over container life will also beevaluated.

v. Particle Size Distribution

Particle size distribution will be evaluated across all batches,formulations as well as under stability testing conditions. Forparticular formulations, the total mass of drug collected on all stagesand accessories will be shown to lie between 85 and 115 percent of labelclaim on a per actuation basis.

vi. Stability studies will be performed on all batches as well as eachformulation. The test storage conditions in the stability protocol for adrug product intended for storage under controlled room temperatureconditions will include (1) accelerated (40±2° C./75±5% RelativeHumidity (RH)), (2) intermediate (30±2° C./60±5% RH), if applicable, and(3) long-term (25±2° C./60±5% RH) conditions.

Examples 8-12

Particle Size Analysis

Particle size analysis was carried out using laser diffraction and lightmicroscopy.

A Malvern Mastersizer 2000 equipped with a Scirocco 2000 SampleDispersion Unit was used to measure the geometric particle size of thesamples. For each measurement approximately 50-100 mg of powder wasloaded into the micro volume tray. Measurement parameters can be foundin Table 24. 3 replicates are run per sample and the average of thethree replicates is reported to 3 decimal places.

TABLE 24 Sampler Scirocco, original configuration Sample Tray MicroVolume Material Name Fraunhofer (RI = 0) Calculation Model GeneralPurpose - normal sensitivity Background 10 seconds Measurement TimeSample Measurement 30 seconds Time Obscuration Limits 1-6% ObscurationFiltering Enable filtering, 30 second time out Feed Rate 75% DispersiveAir 4.0 Bar PressureMorphologyScanning Electron Microscopy

Field emission scanning electron microscopy (FE-SEM, FEI, Sirion, USA)was used to examine the morphology and surface appearance of various ASAparticles. The samples were attached to specimen stubs with two-sidedadhesive tape and Pt-coated with a sputter coater (BAL-TEC, SCD 005,Germany) at 30 mA for 150 s. The coated microcapsules were examinedusing a Sirion SEM at 10 kV with a 1.5 nm resolution according to apreviously reported method (Rosenberg et al., 1985).

HPLC Analysis:

Equipment

The HPLC column was Phenomenex Luna 3u C18(2) 50 mm, 4.6 μm, whichcaused the drug to elute at −1.3 minutes.

HPLC Method Conditions

Analytical Column Phenomenex Luna 3μ C18, 4.0 × 300 mm, 5 μm MobilePhase A 10 mM Sodium Heptanesulfonte in 85:15 Water:Acetonitrile (pH 3.4with Acetic Acid) Mobile Phase B NA Diluent 1% Formic Acid inAcetonitrile Flow Rate 2.0 mL/min Column Temperature 40° C. InjectionVolume 10 μL UV Wavelength 280 nm Run Time 5.0 min (Isocratic)

The particle size measurement is taken by quantifying deposition amountsby HPLC and entering these values into a program CITDAS—Copley InhalerTesting Data Analysis Software.

Batches of aspirin formulation, manufactured using either a jet milledor solution based approach were evaluated for the generalcharacteristics, as well as for the stability studies (see Example 8).For some of the aspirin (ASA) formulations disclosed herein, ASA was jetmilled to <5 μm and suspended at 2 wt % in hexane, followed by spraydrying. Additionally, the impact of including lecithin into theformulation was also tested. Thus, properties of jet milled ASA, whichwas suspended at 2 wt % in hexane, and then spray dried in the presenceor absence of lecithin were evaluated in Example 8.

General manufacturing and yield characteristics of jet-milled control(BREC1511-024, 100% j et-milled ASA), spray dried from hexane, 100% ASA(BREC1511-038A), and spray dried from hexane 99.9/0.1 ASA/Lecithin(BREC1511-038B) are depicted in Table 25 (Example 8).

Next, the accelerated stability study of the formulations was carriedout. Bulk stability, particle size stability, and aerosol stability ofASA formulations are described in Example 8.

Bulk powder aliquots were prepared under dry conditions and placed inamber glass jar, after which they were sealed with desiccant in Mylarbags. The bulk stability study incorporated a 4-week analysis of samplesat 30° C. and 65% relative humidity (RH).

The following formulations were prepared according to the protocoldescribed in Example 8: 100% j et-milled ASA (BREC1511-024),BREC1511-038A (spray dried from hexane containing 100% ASA), and spraydried from hexane containing 99.9/0.1 ASA/Lecithin (BREC1511-038B).

RP-HPLC assay showed that there was no significant loss of potency orincrease in degredents over the period of 4 weeks (Table 26). Also,under the 65% relative humidity conditions, none of the formulationscontained any measurable water, nor was there any uptake detected.

Particle size stability over the period of 4 weeks for formulationsgenerated from milling (FIG. 8) was evaluated. Particle size analysis ateach step (0 week, 1 week, 2 weeks, and 4 weeks) was carried out usingMalvern particle size analyzer.

Of the formulations tested, spray dried milled ASA from 100% hexane(BREC-1511-038A) and spray dried milled ASA suspended in hexane withlecithin (BREC-1511-038B) exhibited particle size stability over 4 weeksof analysis (FIG. 8).

The profile of particles obtained at time 0 (30° C. and 65% relativehumidity (RH)) for spray dried milled ASA suspended in 100% hexane(BREC-1511-038A) was the following: D(v0.1)=0.9 μm; D (v0.5)=2.3 μm; D(v0.9)=4.5 μm; D [3,2]=1.6 μm; and D[4,3]=2.5 μm (FIG. 8). The profileof particles obtained at 4 weeks (30° C. and 65% relative humidity (RH))for spray dried milled ASA suspended in 100% hexane (BREC-1511-038A) wasthe following: D(v0.1)=0.9 μm; D (v0.5)=2.3 μm; D (v0.9)=4.6 μm; D[3,2]=1.7 μm; and D[4,3]=2.6 μm (FIG. 8).

The profile of particles obtained at time 0 (30° C. and 65% relativehumidity (RH)) for spray dried milled ASA suspended in hexane withlecithin (BREC-1511-038B) was the following: D(v0.1)=0.9 μm; D(v0.5)=2.0 μm; D (v0.9)=3.9 μm; D [3,2]=1.6 μm; and D[4,3]=2.2 μm (FIG.8). The profile of particles obtained at 4 weeks (30° C. and 65%relative humidity (RH)) for spray dried milled ASA suspended in hexanewith lecithin (BREC-1511-038B) was the following: D(v0.1)=1.0 μm; D(v0.5)=2.4 μm; D (v0.9)=4.6 μm; D [3,2]=1.7 min; and D[4,3]=2.6 μm (FIG.8).

Taken together, particle size stability studies provide that spray driedmilled ASA suspended in 100% hexane, as well as spray dried milled ASAsuspended in hexane with lecithin, displays stability over the prolongedperiod (about at 30° C. and 65% relative humidity (RH), where theparticle size distribution changed less than 10% over time.

Following the particle size stability analysis, aerosol performanceassay was carried out using a low resistance dry powder inhaler device.The aerodynamic particle size distributions of BREC1511-024,BREC1511-038A, and BREC1511-038B emitted from the dry powder inhaler(DPI) were measured with an eight stage next generation pharmaceuticalimpactor (NGI). The NGI is a particle-classifying cascade impactor fortesting metered-dose, dry-powder, and similar inhaler devices. Oneunique feature of NGI is a micro-orifice collector (MOC) that capturesin a collection cup extremely small particles normally collected on thefinal filter in other impactors. The particles captured in the MOC cupcan be analyzed in the same manner as the particles collected in theother impactor stage cups (Marple et al. Journal of Aerosol Medicine, v.16, (2003).

Particle size distributions at week 0 and week 4 were compareddetermining mass median aerodynamic diameter (MMAD), geometric standarddeviation (GSD), emitted fraction (EF), fine particle fraction (FPF)<5micrometers, and fine particle dose (FPD). Table 27 provides a summaryof findings of aerodynamic properties for each formulation, whereas FIG.9 (BREC1511-024), FIG. 10 (BREC1511-038A), and FIG. 11 (BREC1511-038B),show detailed particle size distribution at week 0 and week 4 based onNGI analysis.

Aerosol performance studies revealed that spray dried milled ASAsuspended in 100% hexane (BREC1511-038A) (FIG. 10), and spray driedmilled ASA suspended in hexane with lecithin (BREC1511-038B) (FIG. 11)maintained the same aerosol properties during the 4 weeks, or did notexhibit a significant aerosol property shift during the 4 weeks. On thecontrary, BREC1511-024, displayed a large shift in properties, withdramatic increase in MMAD and decrease in FPF (FIG. 9).

In conclusion, the results obtained in these studies demonstrate that(1) general potency and purity of bulk powder is unchanged after 4 weeksat 30° C. and 65% RH; (2) spray dried milled ASA suspended in 100%hexane (BREC1511-038A) and spray dried milled ASA suspended in hexanewith lecithin (BREC1511-038B) showed stability over the period of 4weeks; and (3) spray dried milled ASA suspended in 100% hexane(BREC1511-038A) and spray dried milled ASA suspended in hexane withlecithin (BREC1511-038B) do not exhibit any change (or exhibit slightchanges) in the aerosol performance after 4 weeks. On the contrary, jetmilled products changed dramatically after 4 week's condition.

As shown in FIG. 10, BREC1511-038A displayed the followingcharacteristics at 0 time point: MMAD: 3.92±0.13 μm; geometric standarddeviation (GSD): 1.67±0.02; emitted fraction (EF): 63.6±12.7%; fineparticle fraction (FPF)<5 μm: 58.8±3.3%; and fine particle dose (FPD):11.6±1.1 mg. After 4 weeks at 30° C. and 65% RH, BREC1511-038A had thefollowing characteristics: MMAD: 3.90±0.08 μm; GSD: 1.64±0.02; EF:75.1±3.2%; FPF: 58.2±2.2%; and FPD: 12.1±0.8 mg.

As shown in FIG. 11, BREC1511-038B displayed the followingcharacteristics at 0 time point: MMAD: 3.36±0.09 μm; geometric standarddeviation (GSD): 1.67±0.02; emitted fraction (EF): 55.2±3.2%; fineparticle fraction (FPF)<5 μm: 70.1±3.0%; and fine particle dose (FPD):12.9±1.5 mg. After 4 weeks at 30° C. and 65% RH, BREC1511-038B had thefollowing characteristics: MMAD: 3.36±0.09 μm; GSD: 1.73±0.03; EF:73.8±2.3%; FPF: 69.4±2.3%; and FPD: 16.9±1.5 mg.

In certain embodiments, the dry particles of the present compositionhave an MMAD which varies less than about 10%, less than about 6%, orless than about 1%, after the composition is stored at 30° C. at 65%relative humidity for about 4 weeks.

Next, particle size distribution analysis was performed to characterizethe formulation before and after the 2-week storage period at 50° C./75%RH. Average D(0.1), D(0.5) and D(0.9), D(3, 2), and D(4,3) values forBREC1511-024, BREC1511-038A, and BREC1511-038B are shown in Table 11.

As illustrated in Table 11 and FIG. 14, all 3 formulations showed anincrease in particle size after the 2-week storage period at 50° C./75%RH. However, BREC-1511-038A exhibited the least variation between week 0and week 2, while BREC-1511-024 and BREC-1511-038B exhibited similarlevels of variation in particle size distribution before and after the2-week storage period. While the particle size of milled ASA spray driedwith lecithin was slightly smaller compared to ASA alone, the change inparticle size from week 0 to week 2 was greater for BREC-1511-038B thenBREC-1511-038A.

The aerodynamic particle size distributions of BREC1511-024,BREC1511-038A, and BREC1511-038B at week 0 and week 2 after storage at50° C./75% RH were compared determining mass median aerodynamic diameter(MMAD), geometric standard deviation (GSD), emitted fraction (EF), andfine particle fraction (FPF)<5 microm. FIGS. 21-23 illustrate depositionprofile of each aspirin formulation before and after the 2-week periodin the next-generation impactor following aerosolization, where yaxis=deposited fraction (% recovered dose)).

While BREC1511-024 (FIG. 15) and BREC-1511-038B (FIG. 17) displayedshifts in aerosol properties after 2 weeks, BREC1511-038A showed thesmallest change of the 3 formulations (FIG. 16).

Thus, under various temperature, time, and humidity conditions, milledASA suspended in hexane and spray dried, as well as milled ASA suspendedin hexane and spray dried with lecithin, exhibits stability.

In one embodiment, ASA is suspended in hexane prior to spray drying. Inanother embodiment, ASA is suspended in heptane prior to spray drying.In a further embodiment, ASA is suspended in heptane or hexane isomer.In yet another embodiment, ASA is suspended in heptane or hexanederivative prior to spray drying.

Example 8

Batches of aspirin formulations were manufactured and generallycharacterized, manufactured using either a jet milled or solution based(wet polishing) approach.

Aspirin (ASA) was jet milled to <5 μm and suspended at 2 wt % in aparticular solvent. Briefly, ASA solutions were prepared by addingaspirin to the appropriate solvent followed by stirring until ahomogeneous solution was obtained. A BUCHI spray dryer model B-290Advanced was used in all experiments. High performance cyclones wereused to collect the dried product. The spray-drying unit was operated inopen cycle, with the aspirator blowing nitrogen at 100% of capacity,corresponding to a flow rate of the dry nitrogen of approximately 40 kgper hour. Before feeding the stock solution, the spray dryer wasstabilized with the particular solvent. During the stabilization period,the solvent flow rate was adjusted in order to give the target outlettemperature. After stabilization of the outlet temperature, the feed ofthe spray dryer was commuted from the solvent to the product solution(inlet temperature was then readjusted to maintain the outlettemperature in the target value). At the end of the stock solution, thefeed was once more commuted to solvent, in order to rinse the feed lineand carry out a controlled shutdown.

In this specific Example, jet milled ASA was suspended at 2 wt % inhexane, and then spray dried in the presence or absence of lecithin. Theproperties of the ASA formulation were evaluated.

In some cases, an excipient is provided to dry powder formulation inorder to coat the active pharmaceutical ingredient, thus “masking” it.Masking can be useful when the unmodified active pharmaceutical isirritating or otherwise unpleasant to the recipient.

Examples of suitable phospholipid excipients include, withoutlimitation, phosphatidylcholines, phosphatidylethanolamines,phosphatidylinositol, phosphatidylserines, sphingomyelin or otherceramides, as well as phospholipid containing oils such as lecithinoils. As mentioned above, in this example the inventors tested theeffects of lecithin addition to hexane ASA suspension.

General manufacturing and yield characteristics of jet-milled control(BREC1511-024 (100% jet-milled ASA), spray dried from hexane 100% ASA(BREC1511-038A), and spray dried from hexane 99.9/0.1 ASA/Lecithin(BREC1511-038B) are depicted in Table 25.

TABLE 25 General characteristics of BREC1511-024, BREC1511-038A, andBREC1511-038B Spray Reference (BREC-1511) -024 -038A -038B PurposeJet-Milled Neat ASA; spray dry spray dry jet milled ASA Control jetmilled ASA Dry Solids Formulation 100% ASA 99.9/0.1 ASA/Lecithin SprayDryer Scale NA BLD-35 Batch Size (g) 150 6.8 Spray Solution NA 2 Wt%Jet-Milled 2 Wt% Jet-Milled ASA ASA in Hexane + Lecithin Nozzle 2-FluidSolution Feed Rate 12 Atomization Pressure 60 Inlet Temp. (° C.) 115Outlet Temp. (° C.) 60 Yield (%) 81 68 65 D₁₀, D₅₀, D₉₀ 0.7, 1.7, 3.40.9, 2.3, 4.5 0.9, 2.0, 3.9

As seen in Table 25, particle size distribution analysis showed that D10was higher for spray dried formulations than for the initial jet milledproduct. Furthermore, D50 and D90 were higher for BREC1511-038A (100%ASA in hexane, without lecithin) compared to BREC1511-038B (spray driedfrom hexane 99.9/0.1 ASA/Lecithin).

Next, the particle size and particle morphology were evaluated infurther detail. Particle size distribution analysis was performed usingMalvern Particle size analyzer (FIG. 7A). As shown in FIG. 7A, thecollected spray dried particle size was larger than the initial milledproduct. Without being bound to a particular theory, it is expected thatthis is likely related to slight agglomeration or fusion of particlesduring the spray drying process. Furthermore, it is known that cycloneefficiency decreases with smaller particle size, which can result inslight bias of collected particles.

Crystal morphology plays an important role in drug processing anddelivery. Here, particle morphology of jet milled control BREC1511-024,100% ASA BREC1511-038A, and 99.9/0.1 ASA/Lecithin BREC1511-038B wasdetermined by scanning electron microscopy. Briefly, field emissionscanning electron microscopy (FE-SEM, FEI, Sirion, USA) was used toexamine the morphology and surface appearance of various ASA particles.The samples were attached to specimen stubs with two-sided adhesive tapeand Pt-coated with a sputter coater (BAL-TEC, SCD 005, Germany) at 30 mAfor 150 s. The coated microcapsules were examined using a Sirion SEM at10 kV with a 1.5 nm resolution according to a previously reported method(Rosenberg et al., 1985).

Thus, this Example shows that in ASA formulations where hexane is usedas a solvent, milled ASA spray dried with lecithin leads to slightlysmaller particle size as compared to 100% ASA in hexane.

Example 9 Accelerated Stability Study

In the next series of studies (Examples 9 and 10), the bulk stability,particle size stability, and aerosol stability of ASA formulationsdescribed in Example 8 were evaluated.

Bulk Stability

Bulk powder aliquots were prepared under dry conditions and placed inamber glass jar, after which they were sealed with desiccant in Mylarbags. The bulk stability study was designed to incorporate a 4-weekanalysis of samples at 30° C. and 65% relative humidity (RH).

The following formulations were prepared according to the protocoldescribed in Example 8: 100% j et-milled ASA (BREC1511-024),BREC1511-038A (spray dried from hexane 100% ASA), and spray dried fromhexane 99.9/0.1 ASA/Lecithin (BREC1511-038B).

RP-HPLC assay showed that there was no significant loss of potency orincrease in degredents over the period of 4 weeks (Table 26). Also,under the 65% relative humidity conditions, none of the formulationscontained any measureable water, nor was there any uptake detected.

Next, the inventors evaluated particle size stability over the period of4 weeks for formulations generated from milling (FIG. 8). Particle sizeanalysis at each step (0 wk, 1 wk, 2 wk, and 4 wk) was carried out usingMalvern particle size analyzer.

The profile of particles obtained at time 0 (30° C. and 65% relativehumidity (RH)) for spray dried milled ASA suspended in 100% hexane(BREC-1511-038A) was the following: D(v0.1)=0.9 μm; D (v0.5)=2.3 μm; D(v0.9)=4.5 μm; D [3,2]=1.6 μm; and D[4,3]=2.5 μm (FIG. 8). The profileof particles obtained at 4 weeks (30° C. and 65% relative humidity (RH))for spray dried milled ASA suspended in 100% hexane (BREC-1511-038A) wasthe following: D(v0.1)=0.9 μm; D (v0.5)=2.3 μm; D (v0.9)=4.6 μm; D[3,2]=1.7 μm; and D[4,3]=2.6 μm (FIG. 8).

The profile of particles obtained at time 0 (30° C. and 65% relativehumidity (RH)) for spray dried milled ASA suspended in hexane withlecithin (BREC-1511-038B) was the following: D(v0.1)=0.9 μm; D(v0.5)=2.0 μm; D (v0.9)=3.9 μm; D [3,2]=1.6 μm; and D[4,3]=2.2 μm (FIG.8). The profile of particles obtained at 4 weeks (30° C. and 65%relative humidity (RH)) for spray dried milled ASA suspended in hexanewith lecithin (BREC-1511-038B) was the following: D(v0.1)=1.0 μm; D(v0.5)=2.4 μm; D (v0.9)=4.6 μm; D [3,2]=1.7 μm; and D[4,3]=2.6 μm (FIG.8).

Taken together, particle size stability studies provide that spray driedmilled ASA suspended in 100% hexane, as well as spray dried milled ASAsuspended in hexane with lecithin, displays stability over the prolongedperiod (about at 30° C. and 65% relative humidity (RH), where theparticle size distribution changed less than 10% over time.

TABLE 26 RP-HPLC Assay 1 Week 2 Week 4 Week Formulation Initial AssayAssay Assay Lot Approach (w/w %) Assay (ASA) (ASA) (ASA) BREC- Jetmilled 100% ASA —  99.9% ± 0.5 102.9% ± 4.0  99.5% ± 0.3 1511-024 BREC-Suspended 100% ASA 101.3% ± 0.9 101.0% ± 0.9 100.5% ± 0.0  99.6% ± 0.11511-038A Milled BREC- ASA in 99.9/0.1 101.1% ± 0.4 100.0% ± 0.4  99.1%± 0.4 100.6% ± 0.2 1511-038B Hexane ASA/LecithinAerosol Performance

Following the particle size stability analysis, aerosol performanceassay was carried out. Briefly, aerosol performance was determined invitro using a low resistance dry powder inhaler device. The aerodynamicparticle size distributions of BREC1511-024, BREC1511-038A, andBREC1511-038B, emitted from the dry powder inhaler (DPI) were measuredwith an eight stage next generation pharmaceutical impactor (NGI). TheNGI is a particle-classifying cascade impactor for testing metered-dose,dry-powder, and similar inhaler devices. One unique feature of NGI is amicro-orifice collector (MOC) that captures in a collection cupextremely small particles normally collected on the final filter inother impactors. The particles captured in the MOC cup can be analyzedin the same manner as the particles collected in the other impactorstage cups (Marple et al. Journal of Aerosol Medicine, v. 16, (2003).

For each compound formulation, a single size 3 HPMC capsule was filledwith 37 mg of formulated aspirin, and loaded into a RS01 low resistancedevice. Material was actuated at 60 L/min for 4 seconds. Threereplicates were performed per lot. Particle size distributions at week 0and week 4 were compared determining mass median aerodynamic diameter(MMAD), geometric standard deviation (GSD), emitted fraction (EF), fineparticle fraction (FPF)<5 microm, and fine particle dose (FPD). Table 27provides a summary of findings of aerodynamic properties for eachformulation, whereas FIG. 9 (BREC1511-024), FIG. 10 (BREC1511-038A),FIG. 11 (BREC1511-038B), show detailed particle size distribution atweek 0 and week 4 based on NGI analysis.

Aerosol performance studies revealed that the only formulation thatmaintained the same aerosol properties during the 4 weeks was spraydried milled ASA suspended in 100% hexane (BREC1511-038A), (FIG. 10). Onthe contrary, BREC1511-024 displayed a large shift in properties, withdramatic increase in MMAD and decrease in FPF (FIG. 9). WhileBREC1511-038B did not exhibit a significant aerosol shift as the oneobserved for BREC1511-024, a small shift was nevertheless detected (FIG.11). It is possible that the small shift is due to inherent variability,but this remains to be tested. Furthermore, an improved EF wasidentified, which resulted in higher FPD.

TABLE 27 Particle Size Stability at 4 weeks Fine Particle Fraction FineEmitted (FPF) Particle Lot Time Fraction <5 um Dose Number FormulationPoint MMAD GSD (EF) (%) (%) (mgA) BREC1511- Jet Milled Initial 3.66 ±0.23 1.77 ± 0.06 74.1 ± 18.3 60.9 ± 6.3 16.5 ± 2.9 024 ASA 4 week 6.00 ±0.10 1.87 ± 0.01 87.4 ± 1.2  22.1 ± 1.4  7.2 ± 0.4 (Control) BREC1511-100% Initial 3.92 ± 0.13 1.63 ± 0.07 63.6 ± 12.7 58.8 ± 3.3 13.7 ± 2.1038A ASA 4 week 3.90 ± 0.08 1.64 ± 0.02 75.1 ± 3.2  58.2 ± 2.2 16.2 ±0.8 (Hexane) BREC1511- 99.9/0.1 Initial 3.36 ± 0.09 1.67 ± 0.02 55.2 ±3.2  70.1 ± 3.0 14.3 ± 1.1 038B ASA/ 4 week 3.36 ± 0.09 1.73 ± 0.03 73.8± 2.3  69.4 ± 2.3 19.0 ± 0.8 lecithin (Hexane)

In conclusion, the results obtained in these studies demonstrate that(1) general potency and purity of bulk powder is unchanged after 4 weeksat 30° C. and 65% RH; (2) spray dried milled ASA suspended in 100%hexane (BREC1511-038A) and spray dried milled ASA suspended in hexanewith lecithin (BREC1511-038B) showed stability over the period of 4weeks; and (3) spray dried milled ASA suspended in 100% hexane(BREC1511-038A) and spray dried milled ASA suspended in hexane withlecithin (BREC1511-038B) do not exhibit any change (or exhibit slightchanges) in the aerosol performance after 4 weeks. On the contrary, jetmilled products changed dramatically after 4 week's condition.

As shown in FIG. 10, BREC1511-038A displayed the followingcharacteristics at 0 time point: MMAD: 3.92±0.13 μm; geometric standarddeviation (GSD): 1.67±0.02; emitted fraction (EF): 63.6±12.7%; fineparticle fraction (FPF)<5 μm: 58.8±3.3%; and fine particle dose (FPD):11.6±1.1 mg. After 4 weeks at 30° C. and 65% RH, BREC1511-038A had thefollowing characteristics: MMAD: 3.90±0.08 μm; GSD: 1.64±0.02; EF:75.1±3.2%; FPF: 58.2±2.2%; and FPD: 12.1±0.8 mg.

As shown in FIG. 11, BREC1511-038B displayed the followingcharacteristics at 0 time point: MMAD: 3.36±0.09 μm; geometric standarddeviation (GSD): 1.67±0.02; emitted fraction (EF): 55.2±3.2%; fineparticle fraction (FPF)<5 μm: 70.1±3.0%; and fine particle dose (FPD):12.9±1.5 mg. After 4 weeks at 30° C. and 65% RH, BREC1511-038B had thefollowing characteristics: MMAD: 3.36±0.09 μm; GSD: 1.73±0.03; EF:73.8±2.3%; FPF: 69.4±2.3%; and FPD: 16.9±1.5 mg.

In certain embodiments, the dry particles of the present compositionhave an MMAD which varies less than about 10%, less than about 6%, orless than about 1%, after the composition is stored at 30° C. at 65%relative humidity for about 4 weeks.

Example 10 Accelerated Stability Study (2 Weeks at 50° C./75% RH)

In this study, 3 different formulations of spray dried aspirin wereevaluated: BREC1511-024 (100% jet-milled ASA), BREC1511-038A (spraydried from hexane 100% ASA), and BREC1511-038B (spray dried from hexane99.9/0.1 ASA/Lecithin).

Briefly, bulk powder aliquots were prepared under dry conditions andplaced in amber glass jar, after which they were sealed with desiccantin Mylar bags. Each sample was then incubated for 2 weeks at 50° C. and75% relative humidity (RH) conditions. Subsequent to the 2-weekincubation period, the following properties of each formulation wereexamined: 1) particle morphology; 2) water content; 3) potency/purity ofthe formulation; 4) particle size distribution; and 5) NGI analysis(single replicate).

Particle Morphology

Particle morphology was determined by scanning electron microscopy (FIG.12). As shown in FIG. 12, some degree of particle fusion was observed inall 3 samples.

Potency/Purity of the Formulation

Aspirin is in many pharmacopoeia which recommends a titrimetric methodand HPLC for its analysis (British Pharmacopoeia. Vol. 1. London: HerMajesty's Stationary Office; 2009. pp. 442-5; The United StatesPharmacopoeia. Vol. 30. Rockville: U.S. Pharmacopoeial Convention Inc;2008. p. 1164; Patel et al. Indian J Pharm Sci. 75(4): 413-419 (2013)).Using the reversed-phase high-performance liquid chromatography(RP-HPLC) method, degradation studies were performed on each compoundformulation after the 2-week storage period. The chromatogram showingthe peak purity of each compound formulation by UV (FIG. 13)demonstrates the lack of significant degradants after 2 weeks.Furthermore, potency of each of the 3 compound formulations is preservedafter 2 weeks of incubation (50° C./75% RH) (Table 28).

TABLE 28 No significant loss of potency is observed after 2 weeks at 50°C./75% RH Formulation 2 Week Assay Lot Approach (w/w %) Initial Assay(ASA) BREC-1511-024 Jet Milled 100% ASA — 100.5% ± 0.5 BREC-1511-Suspend milled 100% ASA 101.3% ± 0.9 100.4% ± 0.8 038A ASA in hexaneBREC-1511- 99.9/0.1 101.1% ± 0.4 100.1% ± 0.7 038B ASA/LecithinParticle Size Distribution

Next, particle size distribution using Malvern particle size analyzerwas performed to characterize the formulation before and after the 2week period. Average D(0.1), D(0.5) and D(0.9), D[3, 2], and D[4,3]values for BREC1511-024, BREC1511-038A, and BREC1511-038B are shown inTable 29.

TABLE 29 Particle Size Distribution D(v 0.1) D(v 0.5) D(v 0.9) D [3, 2]D [4, 3] Lot μm μm μm μm μm Span BREC-1511-024 0.6 1.5 3.0 1.2 1.7 1.597(T = 0 wk) BREC-1511-024 0.8 3.4 9.0 1.9 4.3 2.401 (T = 2 wk)BREC-1511-038A 0.9 2.3 4.5 1.6 2.5 1.592 (T = 0 wk) BREC-1511-038A 1.12.9 6.0 2.0 3.3 1.720 (T = 2 wk) BREC-1511-038B 0.9 2.0 3.9 1.6 2.21.487 (T = 0 wk) BREC-1511-038B 1.0 3.8 9.2 2.1 4.6 2.171 (T = 2 wk)

As illustrated in Table 29 and FIG. 14, all 3 compound formulationsshowed an increase in particle size after the 2-week storage period.While the particle size of milled ASA spray dried with lecithin wasslightly smaller compared to ASA alone, the change in particle size fromweek 0 to week 2 was greater for BREC-1511-038B then BREC-1511-038A.

Aerosol Performance

The aerodynamic particle size distributions of BREC1511-024,BREC1511-038A, and BREC1511-038B were determined according to thefollowing: for each compound formulation, a single capsule was filledwith 37 mg of formulated aspirin, and loaded into a low resistancedevice. Material was actuated at 60 L/min for 4 seconds. Particle sizedistributions at week 0 and week 2 were compared determining mass medianaerodynamic diameter (MMAD), geometric standard deviation (GSD), emittedfraction (EF), and fine particle fraction (FPF)<5 microm. FIGS. 21-23illustrate deposition profile of each aspirin formulation before andafter the 2-week period in the next-generation impactor followingaerosolization, where y axis=deposited fraction (% recovered dose)).

While BREC1511-024 (FIG. 15) and BREC-1511-038B (FIG. 17) displayedshifts in aerosol properties after 2 weeks, BREC1511-038A showed thesmallest change of the 3 formulations (FIG. 16).

Example 11 Process Optimization

In the next set of experiments, the optimization of suspension spraydrying process was conducted, with the goal of evaluating options forincreasing throughput of hexane formulations by increasing suspensionflow rate and/or increasing suspension concentration. 5 batches ofspray-dried ASA suspension were manufactured on a PSD-1 scale spraydryer at batch sizes up to 300 g using jet-milled ASA from a third partysupplier. The following variables were evaluated: increased spray dryingsuspension feed rate and increased solids concentration in suspension.These included the following formulations: Spray Dried 100% Jet MilledASA (BREC-1511-052A), Spray Dried 100% Jet Milled ASA (HighFlow)(BREC-1511-052B), Spray Dried 100% Jet Milled ASA (High Flow, HighSolids)(BREC-1511-052C), Spray Dried 100% Jet Milled ASA (High Flow,High Solids, High Tout)(BREC-1511-052D), and Spray Dried 99.9/0.1 JetMilled ASA/Lecithin (High Flow)(BREC-1511-052E). The goal was toevaluate the feasibility of spray drying batches with higher solids(BREC-1511-052C) and flow rate (BREC-1511-052B). Table 30 provides asummary of manufacturing characteristics. As shown in FIG. 18, allprocessing conditions produced particles of similar size and within theinhalable range. Furthermore, particle morphology was similar for allfive batches (FIG. 19).

Further characterization of powder demonstrated that all samples havesimilar properties. No measurable amount of residual hexane or water wasfound in powder. Moreover, potency was within the expected range (FIG.20).

Finally, time course of stability of each of the five batches describedin this example at 5 days and 50° C. was studied. The summary of thesefindings is depicted in Table 31. Each of the five batches exhibitedexcellent stability under these conditions.

TABLE 30 Manufacturing Summary -052B Spray Reference -052A (High Liquid-052C -052D -052E (BREC-1511) (Control) Flow Rate) (High Solids) (HighTout) (Lecithin “control” Formulation 109% ASA 99.9/0.1 ASA/LecithinAnti-Solvent Hexane Solid Content 2  2 15 15  2 Batch Size  300 Nozzle1650/120 LC/AC 2850/120 LC/AC Atomization Pressure 40 50 [psi] Flowrate[g/min] 30 150  150  150  150  Drying Gas [g/min] 1250 Inlet Temperature100  155  155  196  145  ° C. Outlet Temperature 60 60 60 78 60 ° C.Yield 58 56 65 74 69

TABLE 31 Particle Size Stability after 5 days (50 C.) Lot Sample StorageD(v 0.1) D(v 0.5) D(v 0.9) D [3, 2] D [4, 3] Number DescriptionCondition μm μm μm μm μm JP Non- 100% Jet Initial 0.7 1.7 3.4 1.3 1.9Refrigerated Milled ASA Lot (Initial) 100% Jet 5 days, 0.8 2.6 5.9 1.73.0 Milled ASA 50° C. (5 days, 50° C.) JP 100% Jet Milled Initial 0.71.9 3.7 1.4 2.1 Refrigerated ASA (Initial) Lot 100% Jet 5 days, 0.8 2.75.8 1.7 3.1 Milled ASA 50° C. (5 days, 50° C.) BREC- Spray Dried Initial0.8 2.0 4.0 1.4 2.2 1511- 100% Jet 052A Milled ASA (Initial, Baseline)Spray Dried 5 days, 0.9 2.6 5.3 1.8 2.9 100% Jet 50° C. Milled ASA (5days, 50° C., Baseline) BREC- Spray Dried Initial 0.9 2.1 3.9 1.7 2.31511- 100% Jet 052B Milled ASA (Initial, High Flow) Spray Dried 5 days,1.0 2.6 5.2 1.8 2.9 100% Jet 50° C. Milled ASA (5 days, 50° C., HighFlow) BREC- Spray Dried Initial 0.8 2.0 3.8 1.5 2.2 1511- 100% Jet 052CMilled ASA (Initial, High Flow, High Solids) Spray Dried 5 days, 0.9 2.55.2 1.8 2.8 100% Jet 50° C. Milled ASA (5 days, 50° C., High Flow, HighSolids) BREC- Spray Dried Initial 0.8 2.2 4.3 1.6 2.4 1511- 100% Jet052D Milled ASA (Initial, High Flow, High Solids, High Tout) Spray Dried5 days, 0.9 2.7 5.4 1.8 3.0 100% Jet 50° C. Milled ASA (5 days, 50° C.,High Flow, High Solids, High Tout) BREC- Spray Dried Initial 0.8 1.9 3.81.5 2.1 1511- 99.9/0.1 Jet 052E Milled ASA/Lecithin (Initial, High Flow)Spray Dried 5 days, 1.0 2.8 5.8 1.9 3.1 99.9/0.1 Jet 50° C. MilledASA/Lecithin (5 days, 50° C., High Flow)

Example 12 Acetylsalicylic Acid Compositions Containing Leucine

First, the solubility of ASA and L-Leucine, as well as the stability ofthe spray solution, was evaluated.

ASA and L-Leucine Absolute Solubility

To define the absolute solubility of ASA and L-leucine in a variety ofsolvent blends, solvent blends were evaluated for solubility of ASA andL-leucine by visual confirmation. As used in this example, unlessspecified otherwise, leucine is L-leucine.

ASA solubility may be up to 5 wt. % total solids in all solvent blendsexcept in 25/75 EtOH/water. Leucine solubility was limited in selectedblends. Possible options for improvement include acidify water fractionto increase L-leucine solubility and decrease salicylic acid conversion(pH 4.0).

TABLE 32 Theoretical Required Solids for Proposed Processing 5 wt. % 2.5wt. % 1 wt. % Total Solids Total Solids Total Solids ASA Leucine ASALeucine ASA Leucine Formulation (wt. %) (wt. %) (wt. %) (wt. %) (wt. %)(wt. %) 95/5  4.75 0.25 2.38 0.12 0.95 0.05 Aspirin/Leucine 85/15 4.250.75 2.12 0.38 0.85 0.15 Aspirin/Leucine

TABLE 33 EtOH/ L-Leucine Water ASA Neutral pH 3 Blend (v/v) mg/ml wt. %mg/ml wt. % mg/ml wt. % 90/10  220.0 21.4 Insoluble NT 75/25  117.6 17.40.2 0.02 50/50  72.5 7.5 0.3 0.03 1.0 0.1 25/75  5.8 0.6 0.4 0.04 NT10/90  NT 9.0 NT 11.0 1.0  0/100 10.0 17.0 1.7

FIG. 21 shows ASA and L-leucine absolute solubility.

ASA and L-leucine Mixing Solubility

The objective is to apply absolute solubility of ASA and L-Leu to amixing study to determine if a spray drying approach is feasible.

ASA and L-leucine was independently dissolved in preferred solvent: ASAin EtOH, and L-leucine in a sulfuric acid solution (pH 4.0) (which mayincrease L-leucine solubility in water).

ASA and L-leucine dissolved very quickly into their respective solvents.

Proposed formulations were evaluated at a final total solids of 7 wt. %and 5 wt. %. In one embodiment, solvent ratio was 25/75 EtOH/Water.Precipitation of components was analyzed visually.

At 7 wt. % solids (85/15 ASA/L-leucine formulation) L-leucineprecipitated out of solution after approximately 60 seconds. Actuallyspray drying mixing step will be <1 minute. At 5 wt. % there was noevidence of precipitation (on the order of minutes) of ASA or L-leucine.

FIG. 22 shows a scheme for studying ASA and L-leucine absolutesolubility.

ASA in Ethanol Spray Solution—Stability Evaluation

ASA degrades to salicylic acid (SA) as a function of time andtemperature due to dissolution. Initial and cold 1 day solutionscontained less than 0.5% SA.

Additional solution stability studies include: stainless vessel, mixed,purged with N2; stainless vessel, unmixed, purged with N2; stainlessvessel, unmixed, 4° C., purged with N2; glass vessel, unmixed, purgedwith N2.

9 wt % ASA was added to ethanol with constant stirring to stainlesssteel vessels. Samples were either stirred at ambient room temperature(approx. 22° C.) or 4° C. and sampled at 1, 3 and 6 days (Table 34).

TABLE 34 Ambient Conditions Controlled 4° C. % ASA % ASA Time Purity %SA Purity % SA Initial 99.9 0.1 99.9 0 1 Day 98.0 1.7 99.1 0.4 3 Day90.5 7.5 97.5 2.0 6 Day 90.5 7.2 96.7 2.3In-Line Mixing Process

It was evaluated whether the addition of L-leucine by an in-line mixingprocess can increase spray-dried ASA product yield and retain or improvechemical/physical properties. FIG. 23 shows a scheme for spray dryprocess optimization.

More than one batch (e.g., up to 2 batches) of approximately 150 gsolids per batch was manufactured on a PSD-1 spray dryer utilizing anin-line mixing process. Formulations included 95%/5% (w/w)ASA/L-leucine, 85%/15% (w/w) ASA/L-leucine, 96%/4% (w/w) ASA/L-leucine,and 87%/13% (w/w) ASA/L-leucine. Details of the processing are in Table35. Similar yields were obtained for both formulations of Table 35.

TABLE 35 Spray Reference BREC-1511-178A BREC-1511-178B Formulation (w/w)96/4 Aspirin/ 87/13 Aspirin/ Leucine Leucine Final Solvent Blend (w/w)70/30 Water/EtOH Solids Content [wt%] 2 Solution Temperature RoomTemperature Nozzle [LC/AC] 1650/120 Atomization Pressure [psi] 85Flowrate [g/min] 40 Drying Gas [g/min] 1250 Inlet Temperature [° C.] 180Outlet Temperature [° C.] 60 Yield (%) 71 79 Residual EtOH Content (wt%)<LOD <LODOne batch of 700 g solids per batch was manufactured on a PSD-1 spraydryer utilizing an in-line mixing process. Formulations included 95%/5%(w/w) ASA/L-leucine, and 85%/15% (w/w). Details of the processing are inTable 35A.

TABLE 35A Spray Reference BREC-1688-036 BREC-1688-046 Formulation (w/w)85/15 Aspirin/ 95/15 Aspirin/Leucine Leucine Batch size (g) 700 SolventBlend (w/w) 60/40 Water/EtOH Solids Content [wt%] 5 Nozzle [LC/AC]1650/120 Atomization Pressure [psi] 40 Flowrate [g/min] 30 Drying Gas[g/min] 1370 Inlet Temperature [° C.] 135 Outlet Temperature [° C.] 60Yield (%) 83 79

In one embodiment, the solids were suspended at 2 wt % in 70/30Water/EtOH or 5% in 60/30 Water/EtOH as applicable, and then spraydried. In another embodiment, ASA was added to ethanol and L-leucine wasadded to the water phase. In one embodiment, sulfuric acid was added inthe 2 wt % suspension ine 70/30 Water/EtOH.

Mixing experiments of ethanol and water were carried out to determineprecipitation of ASA and L-leucine.

Varying solvent ratios were screened and mixing was monitored visually.Solution stability was evaluated at various time points, e.g., 24 hoursafter ASA was dissolved in ethanol.

ASA and L-leucine may re-crystallize while spray-drying.

The properties of the ASA formulation were evaluated, includinggeometric particle size distribution (e.g., using laser diffraction suchas the Malvern Dry Method); particle morphology (e.g., using scanningelectron microscopy (SEM)); ID/Assay and purity (e.g., by HPLC); X-raypowder diffraction (XRPD); and aerosol properties by NGI.

Particle Size Distribution and Particle Morphology

FIGS. 24A-C and Table 36 show laser diffraction data (particle sizedistribution) of 96/4 Aspirin/Leucine (BREC-1511-178A) and 87/13Aspirin/Leucine (BREC-1511-178B) formulations before and after storageat a condition of 4° C., 25° C./60% RH, or 40° C./75% RH for 2 months.Similar particle size was observed between formulations, before andafter exposed to the storage condition as shown by D(v 0.9) (FIGS.24D-E).

FIGS. 24F-I and Table 36 show laser diffraction data (particle sizedistribution) of 85/15 Aspirin/Leucine (BREC-1688-036) and 95/5Aspirin/Leucine (BREC-1688-046) formulations before and after storage ata condition of 25° C./60% RH, 30° C./65% RH, or 40° C./75% RN for 1month or 6 months. Similar particle size was observed betweenformulations, before and after exposed to the storage condition as shownby D(v 0.9) (FIGS. 24J-K).

FIG. 25A shows particle morphology of 96/4 Aspirin/Leucine(BREC-1511-178A) and 87/13 Aspirin/Leucine (BREC-1511-178B)formulations. The images were obtained using SEM (scanning electronmicroscopy), and showed rod-like crystals with small and rough spheres.Particle size and morphology were consistent between lots.

Similarly, FIG. 25B-C shows particle morphology of 96/4 Aspirin/Leucine(BREC-1511-178A) and 87/13 Aspirin/Leucine (BREC-1511-178B) formulationsbefore and after storage at a condition of 4° C., 25° C./60% RH, or 40°C./75% RH for 2 months; FIG. 25D-E shows particle morphology of 85/15Aspirin/Leucine (BREC-1688-036) and 95/5 Aspirin/Leucine (BREC-1688-046)formulations before and after storage at a condition of 25° C./60% RH,30° C./65% RH, or 40° C./75% RN for 1 month; and FIG. 25F-G showsparticle morphology of 85/15 Aspirin/Leucine (BREC-1688-036) and 95/5Aspirin/Leucine (BREC-1688-046) formulations before and after storage ata condition of 25° C./60% RH, 30° C./65% RH, or 40° C./75% RN for 6months. Particle size and morphology showed no significant changesbefore and after exposed to the storage conditions.

TABLE 36 Particle size distribution characterized by laser diffractionfor formulations of various aspirin/leucine content ratio and before andafter stored under various conditions Lot Storage Time D(v 0.1) D(v 0.5)D(v 0.9) D [3, 2] D [4, 3] (Formulation) Condition point μm μm μm μm μmSpan BREC- N/A Initial 0.8 1.9 3.6 1.5 2.1 1.506 1511-178A 4° C. 2 month0.8 1.9 3.6 1.5 2.1 1.472 (96/4 25° C./ 2 month 0.8 1.9 3.6 1.5 2.11.470 Aspirin/ 60% RH Leucine) 40° C./ 2 month 0.8 1.9 3.8 1.5 2.1 1.52475% RH BREC- N/A Initial 0.8 1.7 3.3 1.4 1.9 1.486 1511-178B 4° C. 2month 0.8 1.7 3.3 1.4 1.9 1.466 (87/13 25° C./ 2 month 0.8 1.7 3.4 1.41.9 1.470 Aspirin/ 60% RH Leucine) 40° C./ 2 month 0.8 1.8 3.3 1.4 1.91.481 75% RH BREC- N/A Initial 0.7 1.8 3.7 1.3 2.0 1.669 1688-036 25°C./ 1 month 0.6 1.8 3.7 1.3 2.0 1.713 (85/15 60% RH 6 month 0.6 1.8 3.71.3 2.0 1.696 Aspirin/ 30° C./ 1 month 0.7 1.8 3.8 1.3 2.1 1.697Leucine) 65% RH 6 month 0.7 1.8 3.7 1.3 2.0 1.649 40° C./ 1 month 0.71.9 3.8 1.3 2.1 1.690 75% RH 6 month 0.7 1.9 3.9 1.4 2.1 1.693 BREC- N/AInitial 0.7 2.1 4.2 1.5 2.3 1.649 1688-046 25° C./ 1 month 0.7 2.1 4.21.4 2.3 1.668 (95/5 60% RH 6 month 0.6 2.0 4.1 1.4 2.2 1.715 Aspirin/30° C./ 1 month 0.7 2.1 4.2 1.4 2.3 1.674 Leucine) 65% RH 6 month 0.72.0 4.1 1.4 2.2 1.721 40° C./ 1 month 0.7 2.1 4.2 1.4 2.3 1.689 75% RH 6month 0.7 2.0 4.2 1.4 2.3 1.726Aerosol Profile by NGI

Spray dried powder (BREC-1511-178A and BREC-1511-178B, 37 mg) was loadedinto a size 3 HPMC capsule and actuated out of a dry powder inhaler(high resistance) device at 56 L/min for 4.3 seconds (4 kPa and 4 L).Analysis was performed in triplicate and summarized in Table 37.

Spray dried powder (BREC-1688-046 and BREC-1688-036, about 40 mg) wasloaded into a size 3 HPMC capsule and actuated out of a dry powderinhaler (medium resistance) device at 80 L/min for 4.3 seconds (4 kPaand 4 L). Analysis was performed 6 times and summarized in Table 37.

TABLE 37 Emitted Faction Fine (EF) Particle Capsule Fraction Amount and(FPF) Formulation loaded Resistance MMAD GSD Device <5 μm Sample (w/w)(mg) (L/min) (μm) (μm) (%) (%) BREC- 96/4  37 56 3.70 ± 0.11 1.78 ± 0.0982.1 ± 4.0 55.6 ± 3.4 1511- Aspirin/L- 178A Leucine BREC- 87/13 37 564.13 ± 0.11 1.60 ± 0.04 81.6 ± 1.1 52.4 ± 3.2 1511- Aspirin/L- 178BLeucine BREC- 95/5  38 80 4.15 1688- Aspirin/L- 046 Leucine BREC- 85/1534 80 5.14 1688- Aspirin/L- 036 Leucine

FIGS. 26A-E show aerosol profile studies by NGI of examples of variousASA/Leucine formulations as disclosed herein.

FIG. 26A shows Aerosol profile studies by NGI of 96/4 Aspirin/Leucine(BREC-1511-178A) and 87/13 Aspirin/Leucine (BREC-1511-178B)formulations.

Aerosol profile studies by NGI of 95/5 Aspirin/Leucine (BREC-1688-046)formulation (FIGS. 26B-C, Table 37A) and 85/15 Aspirin/Leucine(BREC-1688-036) formulation (FIGS. 26B-C, Table 37B) showed desiredaerodynamic particle size distribution (APSD).

TABLE 37A Aerosol profile of 95/5 Aspirin/Leucine (BREC-1688-046)formulation % % % % % % Mean NGI1 mass NGI2 mass NGI3 mass NGI4 massNGI5 mass NGI6 mass Mean % Throat 7.71410  27% 6.35491  23% 6.90861  24%6.93904  26% 6.86320  25% 5.29987  24% 6.68  25% Stage 1 5.79545  20%5.30423  19% 8.10888  28% 5.77656  22% 4.08575  15% 5.64953  25% 5.79 22% Stage 2 8.22405  29% 8.98963  33% 7.92446  28% 8.07761  31% 9.52022 35% 6.67581  30% 8.24  31% Stage 3 3.68884  13% 3.89285  14% 3.22571 11% 3.27781  12% 4.03638  15% 2.73028  12% 3.48  13% Stage 4 1.77855  6% 1.74130   6% 1.51993   5% 1.46278   6% 1.73986   6% 1.22168   5%1.58   6% Stage 5 0.61353   2% 0.53024   2% 0.46466   2% 0.48070   2%0.54174   2% 0.39445   2% 0.50   2% Stage 6 0.26559   1% 0.21020   1%0.20206   1% 0.18947   1% 0.23145   1% 0.18649   1% 0.21   1% Stage 70.16852   1% 0.13540   0% 0.12511   0% 0.11646   0% 0.14093   1% 0.11557  1% 0.13   1% Stage 8 0.08858   0% 0.07795   0% 0.06902   0% 0.09927  0% 0.07545   0% 0.05478   0% 0.08   0% Impact 28.34  91% 27.24  92%28.55  92% 26.42  91% 27.23  91% 22.33  91% 26.68  91% or Sum device2.55759   8% 2.27338   8% 2.14849   7% 2.29506   8% 2.32176   8% 2.04546  8% 2.27   8% capsule 0.33856   1% 0.20239   1% 0.38007   1% 0.24517  1% 0.22139   1% 0.21063   1% 0.27   1% Total 31.23 100% 29.71 100%31.08 100% 28.96 100% 29.78 100% 24.58 100% 29.22 100% FPD 10.3472810.72444 9.00844 9.23192 11.31602 7.62862 9.71 FPF 36.51482 39.3879831.55493 34.94330 41.54956 34.16547 36.35 (%) MMAD 4.98789 4.913435.68444 5.17886 4.68920 5.38635 5.14 (μm) GSD 1.93230 1.83685 1.950761.86832 1.78594 1.90329 1.88 FPD/ 30.43 31.54 26.50 27.15 33.28 22.4428.56 nominal dose (%)

TABLE 37B Aerosol profile of 85/15 Aspirin/Leucine (BREC-1688-036)formulation % % % % % % Mean NGI1 mass NGI2 mass NGI3 mass NGI4 massNGI5 mass NGI6 mass Mean % Throat 9.59868  30% 10.30501  35% 8.81892 29% 9.91136  32% 9.35064  30% 9.46780  31% 9.58  31% Stage 1 3.47251 11% 3.36805  11% 3.00100  10% 4.13107  13% 3.64162  12% 3.27259  11%3.48  11% Stage 2 8.79188  28% 7.72229  26% 7.42676  25% 8.63021  28%8.68865  28% 8.32730  27% 8.26  27% Stage 3 5.20870  17% 4.45838  15%5.35574  18% 4.65986  15% 4.92383  16% 5.08799  17% 4.95  16% Stage 42.37035   8% 2.17457   7% 2.82377   9% 2.22028   7% 2.34749   8% 2.46379  8% 2.40   8% Stage 5 0.88669   3% 0.77332   3% 1.13100   4% 0.71628  2% 0.79683   3% 0.82592   3% 0.86   3% Stage 6 0.59525   2% 0.45956  2% 0.63242   2% 0.40816   1% 0.47587   2% 0.47717   2% 0.51   2% Stage7 0.38267   1% 0.29810   1% 0.39299   1% 0.26658   1% 0.29199   1%0.29329   1% 0.32   1% Stage 8 0.24903   1% 0.17255   1% 0.31909   1%0.16792   1% 0.19682   1% 0.16349   1% 0.21   1% Impactor 31.56  93%29.73  90% 29.90  90% 31.11  92% 30.71  92% 30.38  91% 30.57  91% Sumdevice 1.99649   6% 2.74755   8% 2.24816   7% 2.21185   7% 2.22988   7%2.42117   7% 2.31   7% capsule 0.55846   2% 0.54042   2% 1.02638   3%0.48605   1% 0.48384   1% 0.55889   2% 0.61   2% Total 34.11 100% 33.02100% 33.18 100% 33.81 100% 33.43 100% 33.36 100% 33.48 100% FPD 14.1490712.19244 14.49717 12.65427 13.37778 13.54149 13.40 FPF 44.83832 41.0080448.48277 40.67363 43.55632 44.57467 43.86 (%) MMAD 4.13984 4.216633.80621 4.39694 4.24188 4.12008 4.15 (μm) GSD 1.66503 2.02583 1.744001.94454 1.98040 1.66714 1.84 FPD/ 37.23 32.09 38.15 33.30 35.20 35.6435.27 nominal dose (%)Potency and Purity by RP-HPLC

TABLE 38 Formulation Potency SA Purity Sample (w/w) (mgA/g) ASA Purity(%) (%) ASA — — 99.76 ± 0.11 0.24 ± 0.11 Standard BREC- 96/4 921 ± 3499.62 ± 0.03 0.38 ± 0.03 1511-178A Aspirin/L- L-Leucine BREC- 87/13 864± 3  99.56 ± 0.05 0.44 ± 0.05 1511-178B Aspirin/ L-LeucinePhysical Properties by XRPD

FIG. 27 shows XRPD analysis of various examples of ASA/Leucineformulations. Particles were composed of crystalline ASA and Leucine.

Chemical Stability of Aspirin in ASA/Leucine FormulationsBREC-1511-178A, BREC-1511-178B, BREC1688-046, and BREC1688-036

Chemical stability of aspirin in the formulations were tested by RP-HPLCbefore and after the formulations were exposed to various storageconditions. All formulations tested showed good chemical stability,although with ASA purity decreased and SA impurity increased as afunction of time temperature (FIGS. 28A-F, Tables 39A).

BREC-1511-178A and BREC-1511-178B formulations showed high stability,and remained over 99% pure after exposed to 4° C., 25° C./60% RH, or 40°C./75% RH for 2 months (FIGS. 28A-B, Table 39A), with sialicylic acid(SA) remained below 1% during the two-month storage time (FIGS. 28C-D).

TABLE 39A Purity of ASA in two month stability test of BREC-1511-178Aand BREC-1511- 178B formulations Time Point 2 Weeks 1 Month 2 MonthSample ID & ASA Purity SA Impurity ASA Purity SA Impurity ASA Purity SAImpurity Composition Condition (%) (%) (%) (%) (%) (%) ASA — 99.76 0.24— — — — Reference Standard BREC1511- Initial 99.62 0.38 — — — — 178A  4°C. 99.71 0.29 99.70 0.30 99.75 0.25 25° C. 99.64 0.36 99.65 0.35 99.780.22 40° C. 99.57 0.43 99.65 0.35 99.77 0.23 BREC1511- Initial 99.560.44 — — — — 178B  4° C. 99.38 0.62 99.43 0.57 99.61 0.39 25° C. 99.380.62 99.47 0.53 99.61 0.39 40° C. 99.24 0.76 99.25 0.75 99.50 0.50

BREC-1688-046 and BREC-1688-036 formulations also showed high stability,and remained over 99% pure after exposed to 25° C./60% RH, 30° C./65% RHor 40° C./75% RH for 1 month (FIGS. 28E-F, Table 39B). ASA puritydecreased further after the formulations were stored at the variousconditions for 6 months, but still maintained a purity of about 97% orhigher, about 97.5% or higher, about 98% or higher, about 98.5% orhigher, about 98.7% or higher, about 98.8% or higher (Table 39B),

TABLE 39B: Purity of ASA in two month stability test of BREC-1688-046and BREC-1688-036 formulations Sample Condition Formulation ASA Purity(%) SA Impurity (%) BREC1688-046 Initial 99.62 ± 0.06 0.38 ± 0.03 1Month, 25° C. 99.65 ± 0.03 0.35 ± 0.03 1 Month, 30° C. 99.61 ± 0.06 0.39± 0.06 1 Month, 40° C. 95/5 ASA/L-Leucine 99.57 ± 0.03 0.43 ± 0.03 6Months, 25° C. 98.86 ± 0.04 0.62 ± 0.02 6 Months, 30° c.C 98.71 ± 0.020.78 ± 0.02 6 Months, 40° c.C 98.37 ± 0.04 1.08 ± 0.04 BREC1688-036Initial 99.34 ± 0.02 0.66 ± 0.01 1 Month, 25° C. 99.00 ± 0.02 1.00 ±0.02 1 Month, 30° C. 98.83 ± 0.02 1.17 ± 0.02 1 Month, 40° C. 85/15ASA/L-Leucine 98.53 ± 0.04 1.47 ± 0.04 6 Months, 25° C. 97.86 ± 0.011.61 ± 0.03 6 Months, 30° C. 97.43 ± 0.51 2.03 ± 0.52 6 Months, 40° C.97.25* 2.24* *Due to equipment error only a single sample was analyzed

Example 13

Composition 1: A dry powder composition comprising dry particles thatcomprise acetylsalicylic acid or a pharmaceutically acceptable saltthereof, wherein the dry particles have a mass median aerodynamicdiameter (MMAD) within a range of about 0.5 μm to about 10 μm, whereinthe composition further comprises one or more amino acids in an amountranging from about 0.1% (w/w) to about 30% (w/w) of the composition.

Composition 2: The dry powder composition according to composition 1,wherein the amino acid is Leucine.

Composition 3: The dry powder composition according to composition 1,wherein the amino acid is in an amount ranging from about 0.1% (w/w) toabout 20% (w/w) of the composition.

Composition 4: The dry powder composition according to composition 3,wherein the amino acid is in an amount ranging from about 2% (w/w) toabout 20% (w/w) of the composition.

Composition 5: The dry powder composition according to composition 4,wherein the amino acid is in an amount ranging from about 4% (w/w) toabout 15% (w/w) of the composition.

Composition 6: The dry powder composition according to composition 5,wherein the amino acid is in an amount of about 5% (w/w) of thecomposition.

Composition 7: The dry powder composition according to composition 5,wherein amino acid is in an amount of about 15% (w/w) of thecomposition.

Composition 8: The dry powder composition according to composition 1,wherein acetylsalicylic acid or a pharmaceutically acceptable saltthereof is in an amount greater than 40% (w/w) of the composition.

Composition 9: The dry powder composition according to composition 1,wherein acetylsalicylic acid or a pharmaceutically acceptable saltthereof is in an amount greater than 50% (w/w) of the composition.

Composition 10: The dry powder composition according to composition 1,wherein the MMAD ranges from about 2.0 to about 5.0 μm.

Composition 11: The dry powder composition according to composition 11,wherein the MMAD ranges from about 3.0 to about 4.0 am.

Composition 12: The dry powder composition according to any one of theprevious compositions, wherein the dry powder composition maintains apurity of acetylsalicylic acid or the pharmaceutically acceptable saltthereof of about 98.5% or higher, about 99% or higher, or about 99.5% ofhigher after storage at 4° C., 25° C./60% RH, 30° C./65% RH, or 40°C./75% RH for one or two months.

Composition 13: The dry powder composition according to any one of theprevious compositions, wherein the dry powder composition maintains apurity of acetylsalicylic acid or the pharmaceutically acceptable saltthereof of about 95.0% or higher, about 96.5% or higher, about 97.0% ofhigher, about 97.5% or higher, about 98% or higher, about 98.5% orhigher, or about 98.8% or higher, after storage at 4° C., 25° C./60% RH,30° C./65% RH, or 40° C./75% RH for six months.

Composition 14: The dry powder composition according to any one of theprevious compositions, wherein the dry powder composition comprisessialic acid (SA) in an amount of about 5.0% or lower, about 4.0% orlower, about 3.0% or lower, about 2.0% or lower, about 1.0% or lower,about 0.05% or lower, after storage at 4° C., 25° C./60% RH, 30° C./65%RH, or 40° C./75% RH for one month, two months, or six months.

Composition 15: The dry powder composition according to any one of theprevious compositions, wherein the morphology of the dry particlesremains consistent after storage at 4° C., 25° C./60% RH, 30° C./65% RH,or 40° C./75% RH for one month, two months, or six months.

Composition 16: The dry powder composition according to composition 15,wherein the particles comprise crystals.

Composition 17: The dry powder composition according to any one of theprevious compositions, wherein the particle size distribution of the dryparticles remains consistent after storage at 4° C., 25° C./60% RH, 30°C./65% RH, or 40° C./75% RH for one month, two months, or six months.

Composition 18: The dry powder composition according to composition 17,wherein one or more parameters selected from the group consisting ofMMAD, D (v 0.1), D (v0.5), D(v0.9), D[3,2], D[4,3] and span of the dryparticles have a change of about 10% or lower, about 5% or lower, orabout 2.5% or lower, after storage at 4° C., 25° C./60% RH, 30° C./65%RH, or 40° C./75% RH for one month, two months, or six months.

System 19: A drug delivery system effective to reduce the risk of athromboembolic event or treat thrombosis, wherein the system comprisesthe dry powder composition of any one of the previous claims, andwherein acetylsalicylic acid is present at a dose ranging from about 5mg to about 40 mg.

System 20: The drug delivery system according to system 19, furthercomprising clopidogrel.

System 21: The drug delivery system according to system 19, furthercomprising another excipient.

System 22: The drug delivery system according to system 21, wherein theexcipient is sodium lauryl sulfate (SLS), lactose, starch, cellulose,sodium citrate, maltodextrin and/or mannitol.

Method 23: A method of treating an ischemic event, reducing the risk ofa thromboembolic event or treating thrombosis, comprising,administrating to a subject in need thereof a therapeutically effectivedose of the dry powder composition of any of the previous claims.

Use 24: A dry powder composition according to any one of compositions1-18 or a drug delivery system of any one of systems 19-22 for use intreating thrombosis or reducing the risk of a thromboembolic event in asubject.

Use 25: Use of a dry powder composition according to any one ofcompositions 1-18 for the manufacture of a medicament for reducing therisk of a thromboembolic event in a subject.

Method 26: The method according to method 23, or the composition or drugdelivery system according to use 24, or the use according to use 25,wherein the thromboembolic event comprises at least one of unstableangina or a myocardial infarction.

Method 27: The method, the composition, the drug delivery system, or theuse according to method 26, wherein the thromboembolic event comprises atransient ischemic attack.

Method 28: The method, the composition, the drug delivery system, or theuse according to method 26, wherein the thromboembolic event comprises astroke.

Method 29: The method, the composition, the drug delivery system, or theuse according to method 26, wherein the thromboembolic event is treatedwithin about 5 minutes of onset of the ischemic event.

Method 30: The method, the composition, the drug delivery system, or theuse according to method 26, wherein the thromboembolic event is treatedwithin about 10 minutes of onset of the ischemic event.

Method 31: The method, the composition, the drug delivery system, or theuse according to method 26, wherein the thromboembolic event is treatedwithin about 15 minutes of onset of the ischemic event.

The foregoing description is provided to enable a person skilled in theart to practice the various configurations described herein. While thesubject technology has been particularly described with reference to thevarious figures and configurations, it should be understood that theseare for illustration purposes only and should not be taken as limitingthe scope of the subject technology.

There may be many other ways to implement the subject technology.Various functions and elements described herein may be partitioneddifferently from those shown without departing from the scope of thesubject technology. Various modifications to these configurations willbe readily apparent to those skilled in the art, and generic principlesdefined herein may be applied to other configurations. Thus, manychanges and modifications may be made to the subject technology, by onehaving ordinary skill in the art, without departing from the scope ofthe subject technology.

It is understood that the specific order or hierarchy of steps in theprocesses disclosed is an illustration of exemplary approaches. Basedupon design preferences, it is understood that the specific order orhierarchy of steps in the processes may be rearranged. Some of the stepsmay be performed simultaneously. The accompanying method claims presentelements of the various steps in a sample order, and are not meant to belimited to the specific order or hierarchy presented.

It is to be understood that, while the subject technology has beendescribed in conjunction with the detailed description, thereof, theforegoing description is intended to illustrate and not limit the scopeof the subject technology. The citation of any references herein is notan admission that such references are prior art to the presentinvention.

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments of the invention described herein. Such equivalents areintended to be encompassed by the following embodiments.

What is claimed is:
 1. A dry powder composition for delivery byinhalation, said dry powder composition comprising dry particles thatcomprise acetylsalicylic acid or a pharmaceutically acceptable saltthereof, wherein the dry particles have a mass median aerodynamicdiameter (MMAD) within a range of about 0.5 μm to about 10 μm, whereinthe composition further comprises leucine in an amount ranging fromabout 5% (w/w) to about 15% (w/w) of the composition.
 2. The dry powdercomposition of claim 1, wherein the leucine is in an amount ranging fromabout 6% (w/w) to about 15% (w/w) of the composition.
 3. The dry powdercomposition of claim 1, wherein the leucine is in an amount ranging fromabout 6% (w/w) to about 14% (w/w) of the composition.
 4. The dry powdercomposition of claim 1, wherein the leucine is in an amount ranging fromabout 7% (w/w) to about 13% (w/w) of the composition.
 5. The dry powdercomposition of claim 1, wherein the leucine is in an amount of about 5%(w/w) of the composition.
 6. The dry powder composition of claim 1,wherein the leucine is in an amount of about 15% (w/w) of thecomposition.
 7. The dry powder composition of claim 1, wherein theacetylsalicylic acid or pharmaceutically acceptable salt thereof is inan amount greater than 40% (w/w) of the composition.
 8. The dry powdercomposition of claim 1, wherein acetylsalicylic acid or apharmaceutically acceptable salt thereof is in an amount greater than50% (w/w) of the composition.
 9. The dry powder composition of claim 1,wherein the MMAD ranges from about 2.0 to about 5.0 μm.
 10. The drypowder composition of claim 1, wherein the MMAD ranges from about 3.0 toabout 4.0 μm.
 11. The dry powder composition of claim 1, wherein the drypowder composition maintains a purity of acetylsalicylic acid or thepharmaceutically acceptable salt thereof of about 98.5% or higher, about99% or higher, or about 99.5% of higher after storage at 4° C., 25°C./60% RH, 30° C./65% RH, or 40° C./75% RH for one or two months. 12.The dry powder composition of claim 1, wherein the dry powdercomposition maintains a purity of acetylsalicylic acid or thepharmaceutically acceptable salt thereof of about 95.0% or higher, about96.5% or higher, about 97.0% of higher, about 97.5% or higher, about 98%or higher, about 98.5% or higher, or about 98.8% or higher, afterstorage at 4° C., 25° C./60% RH, 30° C./65% RH, or 40° C./75% RH for sixmonths.
 13. The dry powder composition of claim 1, wherein the drypowder composition comprises sialic acid (SA) in an amount of about 5.0%or lower, about 4.0% or lower, about 3.0 or lower, about 2.0% or lower,about 1.0% or lower, about 0.05% or lower, after storage at 4° C., 25°C./60% RH, 30° C./65% RH, or 40° C./75% RH for one month, two months, orsix months.
 14. The dry powder composition of claim 1, wherein themorphology of the dry particles remains consistent after storage at 4°C., 25° C./60% RH, 30° C./65% RH, or 40° C./75% RH for one month, twomonths, or six months.
 15. The dry powder composition of claim 1,wherein the particles comprise crystals.
 16. The dry powder compositionof claim 1, wherein a particle size distribution of the dry particlesremains consistent after storage at 4° C., 25° C./60% RH, 30° C./65% RH,or 40° C./75% RH for one month, two months, or six months.
 17. The drypowder composition of claim 16, wherein one or more parameters selectedfrom the group consisting of MMAD, D (v0.1), D (v0.5), D(v0.9), D[3,2],D[4,3] and span of the dry particles have a change of about 10% orlower, about 5% or lower, or about 2.5% or lower, after storage at 4°C., 25° C./60% RH, 30° C./65% RH, or 40° C./75% RH for one month, twomonths, or six months.
 18. A drug delivery system effective to reducethe risk of a thrombotic event or treat thrombosis, wherein the systemcomprises the dry powder composition of claim 1, and whereinacetylsalicylic acid is present at a dose ranging from about 5 mg toabout 40 mg.
 19. The drug delivery system of claim 18, furthercomprising clopidogrel.
 20. The drug delivery system of claim 18,further comprising another excipient.